DNA sequences from staphylococus aureus bacteriophages 3A, 77, and 96 that encode anti-microbial polypeptides

ABSTRACT

The disclosure concerns particular bacteriophage open reading frames, and portions and products of those open reading frames which have antimicrobial activity. Methods of using such products are also described.

RELATED APPLICATIONS

This application is a continuation in part of U.S. application Ser. No. 09/407,804 filed Sep. 28, 1999, entitled DNA SEQUENCES FROM STAPHYLOCCUS AUREUS BACTERIOPHAGE 77 THAT ENCODE ANTI-MICROBIAL POLYPEPTIDES, which claims the benefit of U.S. Provisional Application No. 60/110,992 filed Dec. 3, 1998 entitled DEVELOPMENT OF NOVEL ANTIMICROBIAL AGENTS BASED ON BACTERIOPHAGE GENOMICS, both of which are hereby incorporated by reference in its entirety, including drawings.

BACKGROUND OF THE INVENTION

This invention relates to the identification of antimicrobial agents and of microbial targets of such agents, and in particular to the isolation of bacteriophage DNA sequences, and their translated protein products, showing anti-microbial activity. The DNA sequences can be expressed in expression vectors. These expression constructs and the proteins produced therefrom can be used for a variety of purposes including therapeutic methods and identification of microbial targets.

The following description is provided to assist the understanding of the reader. None of the information provided or references cited is admitted to be prior art to the present invention.

The frequency and spectrum of antibiotic-resistant infections have, in recent years, increased in both the hospital and community. Certain infections have become essentially untreatable and are growing to epidemic proportions in the developing world as well as in institutional settings in the developed world. The staggering spread of antibiotic resistance in pathogenic bacteria has been attributed to microbial genetic characteristics, widespread use of antibiotic drugs and changes in society that enhance the transmission of drug-resistant organisms (for a review, see Cohen, 1992). This spread of drug resistant microbes is leading to ever-increasing morbidity, mortality and health-care costs.

There are over 160 antibiotics currently available for treatment of microbial infections, all based on a few basic chemical structures and targeting a small number of metabolic pathways: bacterial cell wall synthesis, protein synthesis, and DNA replication. Despite all these antibiotics, a person could succumb to an infection as a result of a resistant bacterial infection. Resistance now reaches all classes of antibiotics currently in use, including: β-lactams, fluoroquinolones, aminoglycosides, macrolide peptides, chloramphenicol, tetracyclines, rifampicin, folate inhibitors, glycopeptides, and mupirocin. There is thus a need for new antibiotics, and this need will not subside given the ability bacteria have to overcome each new agent synthesized. It is also likely that targeting new pathways will play an important role in discovery of these new antibiotics. In fact, a number of crucial cellular pathways, such as secretion, cell division, and many metabolic functions, remain untargeted today.

Most major pharmaceutical companies have on-going drug discovery programs for novel anti-microbials. These are based on screens for small molecule inhibitors (e.g., natural products, bacterial culture media, libraries of small molecules, combinatorial chemistry) of crucial metabolic pathways of the micro-organism of interest. The screening process is largely for cytotoxic compounds and in most cases is not based on a known mechanism of action of the compounds. Classical drug screening programs are being exhausted and many of these pharmaceutical companies are looking towards rational drug design programs. Several small to mid-size biotechnology companies, as well as large pharmaceutical companies, have developed systematic high-throughput sequencing programs to decipher the genetic code of specific micro-organisms of interest. The goal is to identify, through sequencing, unique biochemical pathways or intermediates that are unique to the microorganism. Knowledge of the function of these bacterial genes, may form the rationale for a drug discovery program based on the mechanism of action of the identified enzymes/proteins. However, one of the most critical steps in this approach is the ascertainment that the identified proteins and biochemical pathways are 1) non-redundant and essential for bacterial survival, and 2) constitute suitable and accessible targets for drug discovery. These two issues are not easily addressed since to date, 18 prokaryotic genomes have been sequenced and 200 sequenced genomes are expected by the year 2000. For a majority of the sequenced genomes, less than 50% of the open reading frames (ORFs) have been linked to a known function. Even with the genome of Escherichia coli (E. coli), the most extensively studied bacterium, less than two-thirds of the annotated protein coding genes showed significant similarity to genes with ascribed functions (Rusterholtz and Pohlschroder, 1999). Thus considerable work must be undertaken to identify appropriate bacterial targets for drug screening.

SUMMARY OF THE INVENTION

The present invention is based on the identification of, and demonstration that, specific DNA sequences of a bacteriophage, when introduced into a host bacterium can kill, or inhibit growth, of the host. Thus, these DNA sequences are anti-microbial agents. Information based on these DNA sequences can be utilized to develop peptide mimetics that can also function as anti-microbials. The identification of the host bacterial proteins, targeted by the anti-microbial bacteriophage DNA sequences, can provide novel targets for drug design and compound screening.

In this regard, the terms “inhibit”, “inhibition”, “inhibitory”, and “inhibitor” all refer to a function of reducing a biological activity or function. Such reduction in activity or function can, for example, be in connection with a cellular component (e.g., an enzyme), or in connection with a cellular process (e.g., synthesis of a particular protein), or in connection with an overall process of a cell (e.g., cell growth). In reference to cell growth, the inhibitory effects may be bactericidal (killing of bacterial cells) or bacteriostatic (i.e., stopping or at least slowing bacterial cell growth). The latter slows or prevents cell growth such that fewer cells of the strain are produced relative to uninhibited cells over a given time period. From a molecular standpoint, such inhibition may equate with a reduction in the level of, or elimination of, the transcription and/or translation of a specific bacterial target(s), or reduction or elimination of activity of a particular target biomolecule.

In a first aspect the invention provides methods for identifying a target for antibacterial agents by identifying the bacterial target(s) of at least one inhibitory gene product, e.g., protein from ORFs 33, 41, 79 of bacteriophage 3A, ORF 1 of bacteriophage 77 and ORFs 48, 78, 100 of bacteriophage 96 or a homologous product. Such identification allows the development of antibacterial agents active on such targets. Preferred embodiments for identifying such targets involve the identification of binding of target and phage ORF products to one another. The target molecule may be a bacterial protein or other bacterial biomolecule, e.g., a nucleotprotein, a nucleic acid, a lipid or lipid-containing molecule, a nucleoside or nucleoside derivative, a polysaccharide or polysaccharide-containing molecule, or a peptidoglycan. The phage ORF products may be subportions of a larger ORF product that also binds the host target. Exemplary approaches are described below in the Detailed Description.

Additionally, the invention provides methods for identifying targets for antibacterial agents by identifying homologs of a Staphylococcus aureus target of a bacteriophage 3A ORF product, for example, ORFs 33, 41 or 79, bacteriophage 77 ORF product, such as for example, ORF 1 or bacteriophage 96 ORF products, such as for example, ORFs 48, 78, or 100 product. Such homologs may be utilized in the various aspects and embodiments described herein.

The term “fragment” refers to a portion of a larger molecule or assembly. For proteins, the term “fragment” refers to a molecule which includes at least 5 contiguous amino acids from the reference polypeptide or protein, preferably at least 6, 8, 10, 12, 15, 20, 30, 50 or more contiguous amino acids. In connection with oligo- or polynucleotides, the term “fragment” refers to a molecule which includes at least 15 contiguous nucleotides from a reference polynucleotide, preferably at least 18, 21, 24, 30, 36, 45, 60, 90, 150, or more contiguous nucleotides. Also in preferred embodiments, the fragment has a length in a range with the minimum as described above and a maximum which is no more than 90% of the length (or contains that percent of the contiguous amino acids or nucleotides) of the larger molecule (e.g., of the specified ORF), in other embodiments, the upper limit is no more than 60, 70, or 80% of the length of the larger molecule.

Stating that an agent or compound is “active on” a particular cellular target, such as the product of a particular gene, means that the target is an important part of a cellular pathway which includes that target and that the agent acts on that pathway. Thus, in some cases the agent may act on a component upstream or downstream of the stated target, including a regulator of that pathway or a component of that pathway. In general, an antibacterial agent is active on an essential cellular function, often on a product of an essential gene.

By “essential”, in connection with a gene or gene product, is meant that the host cannot survive without, or is significantly growth compromised, in the absence or depletion of functional product. An “essential gene” is thus one that encodes a product that is beneficial, or preferably necessary, for cellular growth in vitro in a medium appropriate for growth of a strain having a wild-type allele corresponding to the particular gene in question. Therefore, if an essential gene is inactivated or inhibited, that cell will grow significantly more slowly or even not at all. Preferably growth of a strain in which such a gene has been inactivated will be less than 20%, more preferably less than 10%, most preferably less than 5% of the growth rate of the wild-type, or not at all, in the growth medium. Preferably, in the absence of activity provided by a product of the gene, the cell will not grow at all or will be non-viable, at least under culture conditions similar to normal in vivo growth conditions. For example, absence of the biological activity of certain enzymes involved in bacterial cell wall synthesis can result in the lysis of cells under normal osmotic conditions, even though protoplasts can be maintained under controlled osmotic conditions. Preferably, but not necessarily, if such a gene is inhibited, e.g., with an antibacterial agent or a phage product, the growth rate of the inhibited bacteria will be less than 50%, more preferably less than 30%, still more preferably less than 20%, and most preferably less than 10% of the growth rate of the uninhibited bacteria. As recognized by those skilled in the art, the degree of growth inhibition will generally depend on the concentration of the inhibitory agent. In the context of the invention, essential genes are generally the preferred targets of antimicrobial agents. Essential genes can encode target molecules directly or can encode a product involved in the production, modification, or maintenance of a target molecule.

A “target” refers to a biomolecule that can be acted on by an exogenous agent, thereby modulating, preferably inhibiting, growth or viability of a cell. In most cases such a target will be a nucleic acid sequence or molecule, or a polypeptide or protein. However, other types of biomolecules can also be targets, e.g., membrane lipids and cell wall structural components.

The term “bacterium” refers to a single bacterial strain, and includes a single cell, and a plurality or population of cells of that strain unless clearly indicated to the contrary. In reference to bacteria or bacteriophage, the term “strain” refers to bacteria or phage having a particular genetic content. The genetic content includes genomic content as well as recombinant vectors. Thus, for example, two otherwise identical bacterial cells would represent different strains if each contained a vector, e.g., a plasmid, with different phage ORF inserts.

In the context of the phage nucleic acid sequences, e.g., gene sequences, of this invention, the terms “homolog” and “homologous” denote nucleotide sequences from different bacteria or phage strains or species or from other types of organisms that have significantly related nucleotide sequences, and consequently significantly related encoded gene products, preferably having related function. Homologous gene sequences or coding sequences have at least 70% sequence identity (as defined by the maximal base match in a computer-generated alignment of two or more nucleic acid sequences) over at least one sequence window of 48 nucleotides (or at least 99, 150, 200, or even the entire ORF or other sequence of interest), more preferably at least 80 or 85%, still more preferably at least 90%, and most preferably at least 95%. The polypeptide products of homologous genes have at least 35% amino acid sequence identity over at least one sequence window of 18 amino acid residues (or 24, 30, 33, 50, 100, or an entire polypeptide), more preferably at least 40%, still more preferably at least 50% or 60%, and most preferably at least 70%, 80%, or 90%. Preferably, the homologous gene product is also a functional homolog, meaning that the homolog will functionally complement one or more biological activities of the product being compared. For nucleotide or amino acid sequence comparisons where a homology is defined by a % sequence identity, the percentage is determined using BLAST programs (with default parameters (Altschul et al., 1997, “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs, Nucleic Acid Res. 25:3389-3402). Any of a variety of algorithms known in the art which provide comparable results can also be used, preferably using default parameters. Performance characteristics for three different algorithms in homology searching is described in Salamov et al., 1999, “Combining sensitive database searches with multiple intermediates to detect distant homologues.” Protein Eng. 12:95-100. Another exemplary program package is the GCG™ package from the University of Wisconsin.

Homologs may also or in addition be characterized by the ability of two complementary nucleic acid strands to hybridize to each other under appropriately stringent conditions. Hybridizations are typically and preferably conducted with probe-length nucleic acid molecules, preferably 20-100 nucleotides in length. Those skilled in the art understand how to estimate and adjust the stringency of hybridization conditions such that sequences having at least a desired level of complementarity will stably hybridize, while those having lower complementarity will not. For examples of hybridization conditions and parameters, see, e.g.,. Maniatis, T. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor University Press, Cold Spring, N.Y.; Ausubel, F. M. et al. (1994) Current Protocols in Molecular Biology. John Wiley & Sons, Secaucus, N.J. Homologs and homologous gene sequences may thus be identified using any nucleic acid sequence of interest, including the phage ORFs and bacterial target genes of the present invention.

A typical hybridization, for example, utilizes, besides the labeled probe of interest, a salt solution such as 6×SSC (NaCl and Sodium Citrate base) to stabilize nucleic acid strand interaction, a mild detergent such as 0.5% SDS, together with other typical additives such as Denhardt's solution and salmon sperm DNA. The solution is added to the immobilized sequence to be probed and incubated at suitable temperatures to preferably permit specific binding while minimizing non-specific binding. The temperature of the incubations and ensuing washes is critical to the success and clarity of the hybridization. Stringent conditions employ relatively higher temperatures, lower salt concentrations, and/or more detergent than do non-stringent conditions. Hybridization temperatures also depend on the length, complementarity level, and nature (i.e., “GC content”) of the sequences to be tested. Typical stringent hybridizations and washes are conducted at temperatures of at least 40° C., while lower stringency hybridizations and washes are typically conducted at 37° C. down to room temperature (˜25° C.). One of ordinary skill in the art is aware that these conditions may vary according to the parameters indicated above, and that certain additives such as formamide and dextran sulphate may also be added to affect the conditions.

By “stringent hybridization conditions” is meant hybridization conditions at least as stringent as the following: hybridization in 50% formamide, 5×SSC, 50 mM NaH₂PO₄, pH 6.8, 0.5% SDS, 0.1 mg/mL sonicated salmon sperm DNA, and 5×Denhart's solution at 42° C. overnight; washing with 2×SSC, 0.1% SDS at 45° C.; and washing with 0.2×SSC, 0.1% SDS at 45° C. In another example, stringent hybridization conditions should not allow for hybridization of two nucleic acids which differ over a stretch of 20 contiguous nucleotides by more than two bases.

Homologous nucleotide sequences will distinguishably hybridize with a reference sequence with up to three mismatches in ten (i.e., at least 70% base match in two sequences of equal length). Preferably, the allowable mismatch level is up to two mismatches in 10, or up to one mismatch in ten, more preferably up to one mismatch in twenty. (Those ratios can, of course, be applied to larger sequences.)

Preferred embodiments involve identification of binding between ORF product and bacterial cellular component that include methods for distinguishing bound molecules, for example, affinity chromatography, immunoprecipitation, crosslinking, and/or genetic screen methods that permit protein:protein interactions to be monitored. One of skill in the art is familiar with these techniques and common materials utilized (see, e.g., Coligan, J. et al. (eds.) (1995) Current Protocols in Protein Science, John Wiley & Sons, Secaucus, N.J.).

Genetic screening for the identification of protein:protein interactions typically involves the co-introduction of both a chimeric bait nucleic acid sequence (here, the phage ORF to be tested) and a chimeric target nucleic acid sequence that, when co-expressed and having affinity for one another in a host cell, stimulate reporter gene expression to indicate the relationship. A “positive” can thus suggest a potential inhibitory effect in bacteria. This is discussed in further detail in the Detailed Description section below. In this way, new bacterial targets can be identified that are inhibited by specific phage ORF products or derivatives, fragments, mimetics, or other molecules.

Other embodiments involve the identification and/or utilization of a target which is mutated at the site of phage 3A, 77 or 96 protein interaction but still functional in the cell by virtue of their host's relatively unresponsive nature in the presence of expression of ORFs previously identified as inhibitory to the non-mutant or wild-type strain. Such mutants have the effect of protecting the host from an inhibition that would otherwise occur by, for example, competing for binding with the phage ORF product and indirectly allow identification of the precise responsible target. The identified target can then be used for, for example, follow-up studies and anti-microbial development. In certain embodiments, rescue and/or protection from inhibition occurs under conditions in which a bacterial target or mutant target is highly expressed. This is performed, for example, through coupling of the sequence with regulatory element promoters, as known in the art, which regulate expression at levels higher than wild-type at, for example, a level sufficiently higher than the inhibitor can be competitively bound to the highly expressed target such that the bacterium is detectably less inhibited.

Identification of the bacterial target can involve identification of a phage-specific site of action. This can involve a newly identified target, or a target where the phage site of action differs from the site of action of a previously known antibacterial agent or inhibitor. For example, phage T7 genes 0.7 and 2.0 target the host RNA polymerase, which is also the cellular target for the antibacterial agent, rifampin. To the extent that a phage product is found to act at a different site than previously described inhibitors, aspects of the present invention can utilize those new, phage-specific sites for identification and use of new agents. The site of action can be identified by techniques known to those skilled in the art, for example, by mutational analysis, binding competition analysis, and/or other appropriate techniques.

Once a bacterial host target or mutant target sequence has been identified, it too can be conveniently sequenced, sequence analyzed (e.g., by computer), and the underlying gene(s), and corresponding translated product(s) further characterized. Preferred embodiments include such analysis and identification. Preferably such a target has not previously been identified as an appropriate target for antibacterial action.

Also in preferred embodiments in which the bacterial target is a polypeptide or nucleic acid molecule, the identification of a bacterial target of a phage ORF product or fragment includes identification of a cellular and/or biochemical function of the bacterial target. As understood by those skilled in the art, this can, for example, include identification of function by identification of homologous polypeptides or nucleic acid molecules having known function, or identification of the presence of known motifs or sequences corresponding to known function. Such identifications can be readily performed using sequence comparison computer software, such as the BLAST programs and similar other programs and sequence and motif databases.

In embodiments involving expression of a phage ORF in a bacterial strain, in preferred embodiments that expression is inducible. By “inducible” is meant that expression is absent or occurs at a low level until the occurrence of an appropriate environmental stimulus provides otherwise. For the present invention such induction is preferably controlled by an artificial environmental change, such as by contacting a bacterial strain population with an inducing compound (i.e., an inducer). However, induction could also occur, for example, in response to build-up of a compound produced by the bacteria in the bacterial culture, e.g., in the medium. As uncontrolled or constitutive expression of inhibitory ORFs can severely compromise bacteria to the point of eradication, such expression is therefore undesirable in many cases because it would prevent effective evaluation of the strain and inhibitor being studied. For example, such uncontrolled expression could prevent any growth of the strain following insertion of a recombinant ORF, thus preventing determination of effective transfection or transformation. A controlled or inducible expression is therefore advantageous and is generally provided through the provision of suitable regulatory elements, e.g., promoter/operator sequences that can be conveniently transcriptionally linked to a coding sequence to be evaluated. In most cases, the vector will also contain sequences suitable for efficient replication of the vector in the same or different host cells and/or sequences allowing selection of cells containing the vector, i.e., “selectable markers.” Further, preferred vectors include convenient primer sequences flanking the cloning region from which PCR and/or sequencing may be performed. In preferred embodiments where the purification of phage product is desired, preferably the bacterium or other cell type does not produce a target for the inhibitory product, or is otherwise resistant to the inhibitory product.

In preferred embodiments, the target of the phage ORF product or fragment is identified from a bacterial animal pathogen, preferably a mammalian pathogen, more preferably a human pathogen, and is preferably a gene or gene product of such a pathogen. Also in preferred embodiments, the target is a gene or gene product, where the sequence of the target is homologous to a gene or gene product from such a pathogen as identified above.

As used herein, the term “mammal” has its usual biological meaning, and particularly includes bovines, swine, dogs, cats, and humans.

Other aspects of the invention provide isolated, purified, or enriched specific phage nucleic acid and amino acid sequences, subsequences, and homologs thereof from or corresponding to ORFs 33, 41 and 79 from bacteriophage 3A, ORF 1 from bacteriophage 77 or ORFs 48, 78 and 100 from bacteriophage 96 (Staphylococcus aureus host bacterium). Such nucleotide sequences are at least 15 nucleotides in length, preferably at least 18, 21, 24, or 27 nucleotides in length, more preferably at least 30, 50, or 90 nucleotides in length. In certain embodiments, longer nucleic acids are preferred, for example those of at least 120, 150, 200, 300, 600, 900 or more nucleotides. Such sequences can, for example, be amplification oligonucleotides (e.g., PCR primers), oligonucleotide probes, sequences encoding a portion or all of a phage-encoded protein, or a fragment or all of a phage-encoded protein. In preferred embodiments, the nucleic acid sequence or amino acid sequence contains a sequence which has a lower length as specified above, and an upper-length limit which is no more than 50, 60, 70, 80, or 90% of the length of the full-length ORF or ORF product. The upper-length limit can also be expressed in terms of the number of base pairs of the ORF (coding region).

As it is recognized that alternate codons will encode the same amino acid for most amino acids due to the degeneracy of the genetic code, the sequences of this aspect includes nucleic acid sequences utilizing such alternate codon usage for one or more codons of a coding sequence. For example, all four nucleic acid sequences GCT, GCC, GCA, and GCG encode the amino acid, alanine. Therefore, if for an amino acid there exists an average of three codons, a polypeptide of 100 amino acids in length will, on average, be encoded by 3¹⁰⁰, or 5×10⁴⁷, nucleic acid sequences. Thus, a nucleic acid sequence can be modified (e.g., a nucleic acid sequence from a phage as specified above) to form a second nucleic acid sequence encoding the same polypeptide as encoded by the first nucleic acid sequence using routine procedures and without undue experimentation. Thus, all possible nucleic acid sequences that encode the amino acid sequences encoded by the phage 3A ORFs 33, 41, and 79, the phage 77 ORF 1 and the phage 96 ORF 48, 78 and 100 as if all were written out in full, taking into account the codon usage, especially that preferred in the host bacterium.

The alternate codon descriptions are available in common textbooks, for example, Stryer, BIOCHEMISTRY 3^(rd) ed., and Lehninger, BIOCHEMISTRY 3^(rd) ed. Codon preference tables for various types of organisms are available in the literature. Because of the number of sequence variations involving alternate codon usage, for the sake of brevity, individual sequences are not separately listed herein. Instead the alternate sequences are described by reference to the natural sequence with replacement of one or more (up to all) of the degenerate codons with alternate codons from the alternate codon table (Table 2), preferably with selection according to preferred codon usage for the normal host organism or a host organism in which a sequence is intended to be expressed. Those skilled in the art also understand how to alter the alternate codons to be used for expression in organisms where certain codons code differently than shown in the “universal” codon table.

For amino acid sequences, sequences contain at least 5 peptide-linked amino acid residues, and preferably at least 6, 7, 10, 15, 20, 30, or 40, amino acids having identical amino acid sequence as the same number of contiguous amino acid residues in a phage 3A ORF 33, 41, or 79, or phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 product. In some cases longer sequences may be preferred, for example, those of at least 50, 70, or 100 amino acids in length. In preferred embodiments, the sequence has bacteria-inhibiting function when expressed or otherwise present in a bacterial cell which is a host for the bacteriophage from which the sequence was derived.

By “isolated” in reference to a nucleic acid is meant that a naturally occurring sequence has been removed from its normal cellular (e.g., chromosomal) environment or is synthesized in a non-natural environment (e.g., artificially synthesized). Thus, the sequence may be in a cell-free solution or placed in a different cellular environment. The term does not imply that the sequence is the only nucleotide chain present, but that it is essentially free (about 90-95% pure at least) of non-nucleotide material naturally associated with it, and thus is distinguished from isolated chromosomes.

The term “enriched” means that the specific DNA or RNA sequence constitutes a significantly higher fraction (2-5 fold) of the total DNA or RNA present in the cells or solution of interest than in normal or diseased cells or in cells from which the sequence was originally taken. This could be caused by a person by preferential reduction in the amount of other DNA or RNA present, or by a preferential increase in the amount of the specific DNA or RNA sequence, or by a combination of the two. However, it should be noted that enriched does not imply that there are no other DNA or RNA sequences present, just that the relative amount of the sequence of interest has been significantly increased.

The term “significant” is used to indicate that the level of increase is useful to the person making such an increase and an increase relative to other nucleic acids of about at least 2-fold, more preferably at least 5- to 10-fold or even more. The term also does not imply that there is no DNA or RNA from other sources. The other source DNA may, for example, comprise DNA from a yeast or bacterial genome, or a cloning vector such as pUC19. This term distinguishes from naturally occurring events, such as viral infection, or tumor type growths, in which the level of one mRNA may be naturally increased relative to other species of mRNA. That is, the term is meant to cover only those situations in which a person has intervened to elevate the proportion of the desired nucleic acid.

It is also advantageous for some purposes that a nucleotide sequence be in purified form. The term “purified” in reference to nucleic acid does not require absolute purity (such as a homogeneous preparation). Instead, it represents an indication that the sequence is relatively more pure than in the natural environment (compared to the natural level, this level should be at least 2-5 fold greater, e.g., in terms of mg/mL). Individual clones isolated from a genomic or cDNA library may be purified to electrophoretic homogeneity. The claimed DNA molecules obtained from these clones could be obtained directly from total DNA or from total RNA. cDNA clones are not naturally occurring, but rather are preferably obtained via manipulation of a partially purified naturally occurring substance (messenger RNA). The construction of a cDNA library from mRNA involves the creation of a synthetic substance (cDNA) and pure individual cDNA clones can be isolated from the synthetic library by clonal selection of the cells carrying the cDNA library. Thus, the process which includes the construction of a cDNA library from mRNA and isolation of distinct cDNA clones yields an approximately 10⁶-fold purification of the native message. Thus, purification of at least one order of magnitude, preferably two or three orders, and more preferably four or five orders of magnitude is expressly contemplated. A genomic library can be used in the same way and yields the same approximate levels of purification.

The terms “isolated”, “enriched”, and “purified” with respect to the nucleic acids, above, may similarly be used to denote the relative purity and abundance of polypeptides (multimers of amino acids joined one to another by α-carboxyl:α-amino group (peptide) bonds). These, too, may be stored in, grown in, screened in, and selected from libraries using biochemical techniques familiar in the art. Such polypeptides may be natural, synthetic or chimeric and may be extracted using any of a variety of methods, such as antibody immunoprecipitation, other “tagging” techniques, conventional chromatography and/or electrophoretic methods. Some of the above utilize the corresponding nucleic acid sequence.

As indicated above, aspects and embodiments of the invention are not limited to entire genes and proteins. The invention also provides and utilizes fragments and portions thereof, preferably those which are “active” in the inhibitory sense described above. Such peptides or oligopeptides and oligo or polynucleotides have preferred lengths as specified above for nucleic acid and amino acid sequences from phage; corresponding recombinant constructs can be made to express the encoded same. Also included are homologous sequences and fragments thereof.

The nucleotide and amino acid sequences identified herein are believed to be correct, however, certain sequences may contain a small percentage of errors, e.g., 1-5%. In the event that any of the sequences have errors, the corrected sequences can be readily provided by one skilled in the art using routine methods. For example, the nucleotide sequences can be confirmed or corrected by obtaining and culturing the relevant phage, and purifying phage genomic nucleic acids. A region or regions of interest can be amplified, e.g., by PCR from the appropriate genomic template, using primers based on the described sequence. The amplified regions can then be sequenced using any of the available methods (e.g., a dideoxy termination method, for example, using commercially available products). This can be done redundantly to provide the corrected sequence or to confirm that the described sequence is correct. Alternatively, a particular sequence or sequences can be identified and isolated as an insert or inserts in a phage genomic library and isolated, amplified, and sequenced by standard methods. Confirmation or correction of a nucleotide sequence for a phage gene provides an amino acid sequence of the encoded product by merely reading off the amino acid sequence according to the normal codon relationships and/or expressed in a standard expression system and the polypeptide product sequenced by standard techniques. The sequences described herein thus provide unique identification of the corresponding genes and other sequences, allowing those sequences to be used in the various aspects of the present invention. Confirmation of a phage ORF encoded amino acid sequence can also be confirmed by constructing a recombinant vector from which the ORF can be expressed in an appropriate host (e.g., E. coli), purified, and sequenced by conventional protein sequencing methods.

In other aspects the invention provides recombinant vectors and cells harboring phage 3A ORF 33, 41, or 79, or phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 or portions thereof, or bacterial target sequences described herein, preferably where the phage or bacterial sequence is inserted in a recombinant vector. As understood by those skilled in the art, vectors may assume different forms, including, for example, plasmids, cosmids, and virus-based vectors. See, e.g., Maniatis, T. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor University Press, Cold Spring, N.Y.; See also, Ausubel, F. M. et al. (eds.) (1994) Current Protocols in Molecular Biology. John Wiley & Sons, Secaucus, N.J.

In preferred embodiments, the vectors will be expression vectors, preferably shuttle vectors that permit cloning, replication, and expression within bacteria. An “expression vector” is one having regulatory nucleotide sequences containing transcriptional and translational regulatory information that controls expression of the nucleotide sequence in a host cell. Preferably the vector is constructed to allow amplification from vector sequences flanking an insert locus. In certain embodiments, the expression vectors may additionally or alternatively support expression, and/or replication in animal, plant and/or yeast cells due to the presence of suitable regulatory sequences, e.g., promoters, enhancers, 3′ stabilizing sequences, primer sequences, etc. In preferred embodiments, the promoters are inducible and specific for the system in which expression is desired, e.g., bacteria, animal, plant, or yeast. The vectors may optionally encode a “tag” sequence or sequences to facilitate protein purification or protein detection. Convenient restriction enzyme cloning sites and suitable selective marker(s) are also optionally included. Such selective markers can be, for example, antibiotic resistance markers or markers which supply an essential nutritive growth factor to an otherwise deficient mutant host, e.g., tryptophan, histidine, or leucine in the Yeast Two-Hybrid systems described below.

The term “recombinant vector” relates to a single- or double-stranded circular nucleic acid molecule that can be transfected into cells and replicated within or independently of a cell genome. A circular double-stranded nucleic acid molecule can be cut and thereby linearized upon treatment with appropriate restriction enzymes. An assortment of nucleic acid vectors, restriction enzymes, and the knowledge of the nucleotide sequences cut by restriction enzymes are readily available to those skilled in the art. A nucleic acid molecule encoding a desired product can be inserted into a vector by cutting the vector with restriction enzymes and ligating the two pieces together. Preferably the vector is an expression vector, e.g., a shuttle expression vector as described above.

By “recombinant cell” is meant a cell possessing introduced or engineered nucleic acid sequences, e.g., as described above. The sequence may be in the form of or part of a vector or may be integrated into the host cell genome. Preferably the cell is a bacterial cell.

In preferred embodiments, the inserted nucleic acid sequence corresponding to at least a portion of a phage 3A ORF 33, 41, and 79, phage 77 ORF 1 and phage 96 ORF48, 78, and 100 gene product has a length as specified for the isolated purified or enriched nucleic acid sequences in an aspect above.

In another aspect, the invention also provides methods for identifying and/or screening compounds “active on” at least one bacterial target of a bacteriophage inhibitor protein or RNA. Preferred embodiments involve contacting bacterial target proteins with a test compound, and determining whether the compound binds to or reduces the level of activity of the bacterial target, e.g., a bacterial protein. Preferably this is done in vivo under approximately physiological conditions. The compounds that can be used may be large or small, synthetic or natural, organic or inorganic, proteinaceous or non-proteinaceous. In preferred embodiments, the compound is a peptidomimetic, as described herein, a bacteriophage inhibitor protein or fragment or derivative thereof, preferably an “activeportion”, or a small molecule. In particular embodiments, the methods include the identification of bacterial targets as described above or otherwise described herein. Preferably the fragment of a bacteriophage inhibitor protein includes less than 80% of an intact bacteriophage inhibitor protein. Preferably, the at least one target includes a plurality of different targets of bacteriophage inhibitor proteins, preferably a plurality of different targets. The plurality of targets can be in or from a plurality of different bacteria, but preferably is from a single bacterial species.

In embodiments involving binding assays, preferably binding is to a fragment or portion of a bacterial target protein, where the fragment includes less than 90%, 80%, 70%, 60%, 50%, 40%, or 30% of an intact bacterial target protein. Preferably, the at least one bacterial target includes a plurality of different targets of bacteriophage inhibitor proteins, preferably a plurality of different targets. The plurality of targets can be in or from a plurality of different bacteria, but preferably is from a single bacterial species.

A “method of screening” refers to a method for evaluating a relevant activity or property of a large plurality of compounds, rather than just one or a few compounds. For example, a method of screening can be used to conveniently test at least 100, more preferably at least 1000, still more preferably at least 10,000, and most preferably at least 100,000 different compounds, or even more.

In the context of this invention, the term “small molecule” refers to compounds having molecular mass of less than 3000 Daltons, preferably less than 2000 or 1500, still more preferably less than 1000, and most preferably less than 600 Daltons. Preferably but not necessarily, a small molecule is not an oligopeptide.

In a related aspect or in preferred embodiments, the invention provides a method of screening for potential antibacterial agents by determining whether any of a plurality of compounds, preferably a plurality of small molecules, is active on at least one target of a bacteriophage inhibitor protein or RNA. Preferred embodiments include those described for the above aspect, including embodiments which involve determining whether one or more test compounds bind to or reduce the level of activity of a bacterial target, and embodiments which utilize a plurality of different targets as described above.

The identification of bacteria-inhibiting phage ORFs and their encoded products also provides a method for identifying an active portion of such an encoded product. This also provides a method for identifying a potential antibacterial agent by identifying such an active portion of a phage ORF or ORF product. In preferred embodiments, the identification of an active portion involves one or more of mutational analysis, deletion analysis, or analysis of fragments of such products. The method can also include determination of a 3-dimensional structure of an active portion, such as by analysis of crystal diffraction patterns. In further embodiments, the method involves constructing or synthesizing a peptidomimetic compound, where the structure of the peptidomimetic compound corresponds to the structure of the active portion.

In this context, “corresponds” means that the peptidomimetic compound structure has sufficient similarities to the structure of the active portion that the peptidomimetic will interact with the same molecule as the phage protein and preferably will elicit at least one cellular response in common which relates to the inhibition of the cell by the phage protein.

The methods for identifying or screening for compounds or agents active on a bacterial target of a phage-encoded inhibitor can also involve identification of a phage-specific site of action on the target.

An “active portion” as used herein denotes an epitope, a catalytic or regulatory domain, or a fragment of a bacteriophage inhibitor protein that is responsible for, or a significant factor in, bacterial target inhibition. The active portion preferably may be removed from its contiguous sequences and, in isolation, still effect inhibition.

By “mimetic” is meant a compound structurally and functionally related to a reference compound that can be natural, synthetic, or chimeric. In terms of the present invention, a “peptidomimetic,” for example, is a compound that mimics the activity-related aspects of the 3-dimensional structure of a peptide or polypeptide in a non-peptide compound, for example mimics the structure of a peptide or active portion of a phage- or bacterial ORF-encoded polypeptide.

A related aspect provides a method for inhibiting a bacterial cell by contacting the bacterial cell with a compound active on a bacterial target of a bacteriophage inhibitor protein or RNA encoded by bacteriophage 3A ORF 33, 41, or 79, bacteriophage 77 ORF 1, or bacteriophage 96 ORF 48, 78, or 100, where the target was uncharacterized. In preferred embodiments, the compound is such a protein, or a fragment or derivative thereof; a structural mimetic, e.g., a peptidomimetic, of such a protein or fragment; a small molecule; the contacting is performed in vitro, the contacting is performed in vivo in an infected or at risk organism, e.g., an animal such as a mammal or bird, for example, a human, or other mammal described herein, or in a plant.

In the context of this invention, the term “bacteriophage inhibitor protein” refers to a protein encoded by a bacteriophage nucleic acid sequence which inhibits bacterial function in a host bacterium. Thus, it is a bacteria-inhibiting phage product.

In the context of this invention, the phrase “contacting the bacterial cell with a compound active on a bacterial target of a bacteriophage inhibitor protein” or equivalent phrases refer to contacting with an isolated, purified, or enriched compound or a composition including such a compound, but specifically does not rely on contacting the bacterial cell with an intact naturally occurring phage which encodes the compound. Preferably no intact phage are involved in the contacting.

Related aspects provide methods for prophylactic or therapeutic treatment of a bacterial infection by administering to an infected, challenged or at risk organism a therapeutically or prophylactically effective amount of a compound active on a target of a bacteriophage 3A ORF 33, 41, or 79, bacteriophage 77 ORF 1, or bacteriophage 96 ORF 48, 78, or 100 product, e.g., as described for the previous aspect. Preferably the bacterium involved in the infection or risk of infection produces the identified target of the bacteriophage inhibitor protein or alternatively produces a homologous target compound. In preferred embodiments, the host organism is a plant or animal, preferably a mammal or bird, and more preferably, a human or other mammal described herein. Preferred embodiments include, without limitation, those as described for the preceding aspect.

Compounds useful for the methods of inhibiting, methods of treating, and pharmaceutical compositions can include novel compounds, but can also include compounds which had previously been identified for a purpose other than inhibition of bacteria. Such compounds can be utilized as described and can be included in pharmaceutical compositions.

By “treatment” or “treating” is meant administering a compound or pharmaceutical composition for prophylactic and/or therapeutic purposes. The term “prophylactic treatment” refers to treating a patient or animal that is not yet infected but is susceptible to or otherwise at risk of a bacterial infection. The term “therapeutic treatment” refers to administering treatment to a patient already suffering from infection.

The term “bacterial infection” refers to the invasion of the host organism, animal or plant, by pathogenic bacteria. This includes the excessive growth of bacteria which are normally present in or on the body of the organism, but more generally, a bacterial infection can be any situation in which the presence of a bacterial population(s) is damaging to a host organism. Thus, for example, an organism suffers from a bacterial infection when excessive numbers of a bacterial population are present in or on the organism's body, or when the effects of the presence of a bacterial population(s) is damaging to the cells, tissue, or organs of the organism.

The terms “administer”, “administering”, and “administration” refer to a method of giving a dosage of a compound or composition, e.g., an antibacterial pharmaceutical composition, to an organism. Where the organism is a mammal, the method is, e.g., topical, oral, intravenous, transdermal, intraperitoneal, intramuscular, or intrathecal. The preferred method of administration can vary depending on various factors, e.g., the components of the pharmaceutical composition, the site of the potential or actual bacterial infection, the bacterium involved, and the infection severity.

The term “mammal” has its usual biological meaning, referring to any organism of the Class Mammalia of higher vertebrates that nourish their young with milk secreted by mammary glands, e.g., mouse, rat, and, in particular, human, bovine, sheep, swine, dog, and cat.

In the context of treating a bacterial infection a “therapeutically effective amount” or “pharmaceutically effective amount” indicates an amount of an antibacterial agent, e.g., as disclosed for this invention, which has a therapeutic effect. This generally refers to the inhibition, to some extent, of the normal cellular functioning of bacterial cells that renders or contributes to bacterial infection.

The dose of antibacterial agent that is useful as a treatment is a “therapeutically effective amount.” Thus, as used herein, a therapeutically effective amount means an amount of an antibacterial agent that produces the desired therapeutic effect as judged by clinical trial results and/or animal models. This amount can be routinely determined by one skilled in the art and will vary depending on several factors, such as the particular bacterial strain involved and the particular antibacterial agent used.

In connection with claims to methods of inhibiting bacteria and therapeutic or prophylactic treatments, “a compound active on a target of a bacteriophage inhibitor protein” or terms of equivalent meaning differ from administration of or contact with an intact phage naturally encoding the full-length inhibitor compound. While an intact phage may conceivably be incorporated in the present methods, the method at least includes the use of an active compound as specified different from a full length inhibitor protein naturally encoded by a bacteriophage and/or a delivery or contacting method different from administration of or contact with an intact phage naturally encoding the full-length protein. Similarly, pharmaceutical compositions described herein at least include an active compound or composition different from a phage naturally coding the full-length inhibitor protein, or such a full-length protein is provided in the composition in a form different from being encoded by an intact phage. Preferably the methods and compositions do not include an intact phage.

In accordance with the above aspects, the invention also provides antibacterial agents and compounds active on a bacterial target of bacteriophage 3A ORF 33, 41, or 79, bacteriophage 77 ORF 1, or bacteriophage 96 ORF 48, 78, or 100, where the target was uncharacterized as indicated above. As previously indicated, such active compounds include both novel compounds and compounds which had previously been identified for a purpose other than inhibition of bacteria. Such previously identified biologically active compounds can be used in embodiments of the above methods of inhibiting and treating. In preferred embodiments, the targets, bacteriophage, and active compound are as described herein for methods of inhibiting and methods of treating. Preferably the agent or compound is formulated in a pharmaceutical composition which includes a pharmaceutically acceptable carrier, excipient, or diluent. In addition, the invention provides agents, compounds, and pharmaceutical compositions where an active compound is active on an uncharacterized phage-specific site on the target.

In preferred embodiments, the target is as described for embodiments of aspects above.

Likewise, the invention provides a method of making an antibacterial agent. The method involves identifying a target of a bacteriophage 3A ORF 33, 41, or 79, bacteriophage 77 ORF 1, or bacteriophage 96 ORF 48, 78, or 100 product, screening a plurality of compounds to identify a compound active on the target, and synthesizing the compound in an amount sufficient to provide a therapeutic effect when administered to an organism infected by a bacterium naturally producing the target.

In preferred embodiments, the identification of the target and identification of active compounds include steps or methods and/or components as described above (or otherwise herein) for such identification. Likewise, the active compound can be as described above, including fragments and derivatives of phage inhibitor proteins, peptidomimetics, and small molecules. As recognized by those skilled in the art, peptides can be synthesized by expression systems and purified, or can be synthesized artificially by methods well known in the art.

In the context of nucleic acid or amino acid sequences of this invention, the term “corresponding” and “correspond” indicates that the sequence is at least 95% identical, preferably at least 97% identical, and more preferably at least 99% identical to a sequence from the specified phage genome or bacterial genome, a ribonucleotide equivalent, a degenerate equivalent (utilizing one or more degenerate codons), or a homologous sequence, where the homolog provides functionally equivalent biological function.

In embodiments where the bacterial target of a bacteriophage inhibitor ORF product, e.g., an inhibitory protein or polypeptide, the target is preferably encoded by a S. aureus nucleic acid coding sequence from a host bacterium for bacteriophages 3A, 77, or 96. Target sequences are described herein by reference to sequence source sites. The sequence encoding the target preferably corresponds to a S. aureus nucleic acid sequence available from numerous sources including S. aureus sequences deposited in GenBank, S. aureus sequences found in European Patent Application No. 97100110.7 to Human Genome Sciences, Inc. filed Jan. 7, 1997, S. aureus sequences available from TIGR at http://www.tigr.org/tdb/mdb/mdb.html, and S. aureus sequences available from the Oklahoma University S. aureus sequencing project at the following URL:

http://www.genome.ou.edu/staph_new.html.

The amino acid sequence of a polypeptide target is readily provided by translating the corresponding coding region. For the sake of brevity, the sequences are not reproduced herein. Also, in preferred embodiments, a target sequence corresponds to a S. aureus coding sequences corresponding to a sequence listed in Table 7. The listings in Table 7 describe S. aureus sequences currently deposited in GenBank. Again, for the sake of brevity, the sequences are described by reference to the GenBank entries instead of being written out in full herein. In cases where an entry for a coding region is not complete, the complete sequence can be readily obtained by routine methods, by isolating a clone in a phages 3A, 77, and 96 host S. aureus genomic library, and sequencing the clone insert to provide the relevant coding region. The boundaries of the coding region can be identified by conventional sequence analysis and/or by expression in a bacterium in which the endogenous copy of the coding region has been inactivated and using subcloning to identify the functional start and stop codons for the coding region.

In an additional aspect, the present invention provides a nucleic acid segment which encodes a protein and corresponds to a segment of the nucleic acid sequence of an ORF (open reading frame) from Staphylococcus aureus bacteriophages 3A, 77 or 96 as provided in Table 1. Preferably, the protein is a functional protein. One of ordinary skill in the art would recognize that bacteriophage possess genes which encode proteins which may be either beneficial or detrimental to a bacterial cell. Such proteins act to replicate DNA, translate RNA, manipulate DNA or RNA, and enable the phage to integrate into the bacterial genome. Proteins from bacteriophage can function as, for example, a polymerase, kinase, phosphatase, helicase, nuclease, topoisomerase, endonuclease, reverse transcriptase, endoribonuclease, dehydrogenase, gyrase, integrase, carboxypeptidase, proteinase, amidase, transcriptional regulators and the like, and/or the protein may be a functional protein such as a chaperon, capsid protein, head and tail proteins, a DNA or RNA binding protein, or a membrane protein, all of which are provided as non-limiting examples. Proteins with functions such as these are useful as tools for the scientific community.

Thus, the present invention provides a group of novel proteins from bacteriophage which can be used as tools for biotechnical applications such as, for example, DNA and/or RNA sequencing, polymerase chain reaction and/or reverse transcriptase PCR, cloning experiments, cleavage of DNA and/or RNA, reporter assays and the like. Preferably, the protein is encoded by an open reading frame in the nucleic acid sequences of bacteriophages 3A, 77 or 96. Within the scope of the present invention are fragments of proteins and/or truncated portions of proteins which have been either engineered through automated protein synthesis, or prepared from nucleic acid segments which correspond to segments of the nucleic acid sequences of bacteriophages 3A, 77 or 96, and which are then inserted into cells via plasmid vectors which can be induced to express the protein. It is understood by one of skill in the art that mutational analysis of proteins has been known to help provide proteins which are more stable and which have higher and/or more specific activities. Such mutations are also within the scope of the present invention, hence, the present invention provides a mutated protein and/or the mutated nucleic acid segment from bacteriophages 3A, 77 or 96 which encodes the protein.

In another aspect, the invention provides antibodies which bind proteins encoded by a nucleic acid segment which corresponds to the nucleic acid sequence of an ORF (open reading frame) from Staphylococcus aureus bacteriophages 3A, 77 or 96 as provided in Table 1. Bacteriophages are bacterial viruses which contain nucleic acid sequences which encode proteins that can correspond to proteins of other bacteriophages and other viruses. Antibodies targeted for proteins encoded by nucleic acid segments of phages 3A, 77 or 96 can serve to bind targets encoded by nucleic acid segments from other viruses which correspond to the sequences provided in Table 1. Furthermore, antibodies to proteins encoded by nucleic acid segments of phages 3A, 77 or 96 can also bind to proteins from other viruses that share similar functions but may not share corresponding sequences. It is understood in the art that proteins with similar activities/functions from a variety of sources generally share motifs, regions, or domains which correspond. Thus, antibodies to motifs, regions, or domains of functional proteins from phages 3A, 77 or 96 should be useful in detecting corresponding proteins in other bacteriophages and viruses. Such antibodies can also be used to detect the presence of a virus sharing a similar protein. Preferably the virus to be detected is pathogenic to a mammal, such as a dog, cat, bovine, sheep, swine, or a human.

As used in the claims to describe the various inventive aspects and embodiments, “comprising” means including, but not limited to, whatever follows the word “comprising”. Thus, use of the term “comprising” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present. By “consisting of” is meant including, and limited to, whatever follows the phrase “consisting of”. Thus, the phrase “consisting of” indicates that the listed elements are required or mandatory and that no other elements may be present. By “consisting essentially of” is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending upon whether or not they affect the activity or action of the listed elements.

Additional features and embodiments of the present invention will be apparent from the following Detailed Description and from the claims, all within the scope of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 are flow schematics showing the manipulations necessary to convert pT0021, an arsenite inducible vector containing the luciferase gene, into a) pTM, b) pTMSM or c) pTHA three ars inducible vectors. Vector pTM contains Bam HI and Hind III cloning sites. Vector pTMSM contains BamHI, SalI and MluI cloning sites. Vector pTHA contains BamHI and SalI cloning sites and a downstream HA epitope tag. This figure also shows in d), the characteristic of the lactose-inducible vector pTMSLac containing Bam HI and SalI cloning sites.

FIG. 2 is a schematic representation of the cloning steps involved to place the DNA segments of any ORFs e.g. 3A ORF 33, 41, 79, or 77 ORF 1, or 96 ORF 48, 78, 100 or other sequences into vector to assess inhibitory potential. For subcloning into a) pTM (and similarly into pTMSM and pTMSLac) individual ORFs were amplified by the PCR using oligonucleotides targetting the start and stop codons of the ORFs. Using this strategy cloning sites (here BamHI and HindIII), were positioned immediately upstream or downstream, respectively of the start and stop codons of each ORF. Following digestion with BamHI and HindIII, the PCR fragments were subcloned into the same sites of pTM (BamHI and HindIII). For subcloning into b) pTHA, individual ORFs e.g. 3A ORF 33, 41, 77 ORF 1 and 96 ORF 48, 78 were amplified by the PCR using oligonucleotides targetting the start codon and the penultimated codon of the ORFs. Using this strategy, BamHI and SalI sites were positioned immediately upstream or downstream, respectively of these two codons. Following digestion with BamHI and SalI, the PCR fragments were subcloned into the same sites of pTHA. Clones were verified by direct sequencing.

FIG. 3 shows a schematic representation of the functional assays used to characterize the bactericidal and bacteriostatic potential of predicted ORFs (>33 amino acids) encoded by bacteriophages 3A, 77, 96. FIG. 3a) Functional assay on semi-solid support media. FIG. 3b) Functional assay in liquid culture.

FIG. 4 shows the results of the functional assay on semi-solid support media to identify bacteriophage 3A, 77 and 96 ORFs with anti-microbial activity. FIG. 4a) shows the lists of the bacteriophage 3A, 77 and 96 ORFs that were screened in the functional assay and FIG. 4b) shows inhibition of bacterial growth following induction of expression of phage 3A ORF 33, 41 and 79, phage 77 ORF1 and phage 96 ORF 48 and 100 from three clones of Staphylococcus aureus transformants. One clone of Staphylococcus aureus transformed with the non-inhibitory ORF (44AHJD bacteriophage ORF 114 cloned into pTM vector) was used as control. From these experiments, it is clear that expression of these ORFs leads to the inhibition of growth of Staphylococcus aureus.

FIG. 5 are the graphs of OD₅₆₅ values and colony forming units (CFU) over time showing the results of functional assay in liquid media to assess bacteriostatic or bactericidal activity of bacteriophage 3A ORF 33, 41 and 79, bacteriophage 77 ORF 1 and bacteriophage 96 ORF 48, 78 and 100. Growth inhibition assays were performed as detailed in the Detailed Description. The OD₅₆₅ values and the number of CFU were determined from cultures of Staphylococcus aureus transformants harboring a given bacteriophage inhibitory ORF, in the absence or presence of the inducer. The identity of the expression vector and subcloned ORF harbored by the Staphylococcus aureus is given at the top of the each graph. The value of OD and the number of CFU was also determined from non-induced and induced control cultures of Staphylococcus aureus transformants harboring a non-inhibitory phage ORF cloned into the same vector. Each graph represents the average obtained from three Staphylococcus aureus transformants.

FIG. 6 shows the pattern of protein expression of the inhibitory ORF in S. aureus in the presence or in the absence of induction with sodium arsenite. Individual inhibitory ORF (phage 3A ORF 33, 41 and 79, phage 77 ORF 1, phage 96 ORF 48, 78 and 100) were subcloned into the pTHA vector. This vector contains BamH I, Sal I cloning sites and a downstream HA epitope tag. The HA tag is set inframe with the ORF and is positioned at the carboxy terminus of each ORF. An anti-HA tag antibody was used for the detection of the ORF expression. The identity of the subcdoned ORF harbored by the Staphylococcus aureus transformants is given at the top of each panel.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Preliminarily the tables will be briefly described.

Table 1 shows the complete nucleotide sequence of the genomes of Staphylococcus aureus bacteriophages 3A, 77 and 96.

Table 2 is a table from Alberts et al., MOLECULAR BIOLOGY OF THE CELL 3^(rd) ed., showing the redundancy of the “universal” genetic code.

Table 3 shows the nucleotide and predicted amino acid sequences of ORF 33, 41, and 79 from bacteriophage 3A, ORF1 from bacteriophage 77, and ORF 48, 78, and 100 from bacteriophage 96.

Table 4 shows the sequence similarities identified to date between ORFs predicted to be encoded by Staphylococcus aureus bacteriophages 3A, 77 and 96 and sequences present in the Genbank and Swissprot sequence databases. More specifically, these results indicate that: I) ORF 1 from phage 77 has significant homology to various genes in the NCBI non-redundant nucleotide database—such as the gene encoding for ORF 16 of the bacteriophage phi PVL, and II) ORF 48 from phage 96 has significant homology to one gene in the NCBI non-redundant nucleotide database—the gene encoding ORF 39 of the bacteriophage phi PVL.

Table 5 shows sequence alignment of phage 96 inhibitory ORFs with other identified inhibitory phage ORFs. The results of this search indicate that the inhibitory 96 ORF 100 has significant homology to the the inhibitory 3A ORF 79 and 96 ORF 48 has a significant homology to the previously identified phage 77 inhibitory ORFs 43 and 182.

Table 6 shows the physiochemical parameters of phage 3A ORF 33, 41, 79, phage 77 ORF 1 and phage 96 ORF 48, 78, 100. These include the primary amino acid sequence of the predicted protein, the average molecular weight, amino acid composition, theoretical pI and hydrophobicity properties.

Table 7 shows S. aureus coding sequences corresponding sequences currently deposited in GenBank.

The present invention is based on the identification of naturally-occurring DNA sequence elements encoding RNA or proteins with anti-microbial activity. Bacteriophages or phages, are viruses that infect and kill bacteria. They are natural enemies of bacteria and, over the course of evolution have perfected enzymes and proteins (products of DNA sequences) which enable them to infect a host bacteria, replicate their genetic material, usurp host metabolism, and ultimately kill their host. The scientific literature documents well the fact that many known bacteria have a large number of such bacteriophages that can infect and kill them (for example, see the ATCC bacteriophage collection at http://www.atcc.org) (Ackermann and DuBow, 1987). Although we know that many bacteriophages encode proteins which can significantly alter their host's metabolism, determination of the killing potential of a given bacteriophage gene product can only be assessed by expressing the gene product in the target bacterial strain.

As indicated in the Summary above, the present invention is concerned with the use of bacteriophage 3A, 77, and 96 coding sequences and the encoded polypeptides or RNA transcripts to identify bacterial targets for potential new antibacterial agents. Thus, the invention concerns the selection of relevant bacteria. Particularly relevant bacteria are those which are pathogens of a complex organism such as an animal, e.g., mammals, reptiles, and birds, and plants. However, the invention can be applied to any bacterium (whether pathogenic or not) for which bacteriophage are available or which are found to have cellular components closely homologous to components targeted by phage 3A ORF 33, 41, 79, phage 77 ORF 1, and phage 96 ORF 48, 78, 100.

Identification of ORFs 33, 41 and 78 from phage 3A, ORF 1 from phage 77 and ORF 48, 78, 100 from phage 96 and products from the phage which inhibit the host bacterium both provides an inhibitor compound and allows identification of the bacterial target affected by the phage-encoded inhibitor. Such a target is thus identified as a potential target for development of other antibacterial agents or inhibitors and the use of those targets to inhibit those bacteria. As indicated above, even if such a target is not initially identified in a particular bacterium, such a target can still be identified if a homologous target is identified in another bacterium. Usually, but not necessarily, such another bacterium would be a genetically closely related bacterium. Indeed, in some cases, an inhibitor encoded by phage 3A ORF 33, 41, or 79, phage 77 ORF1 or phage 96 ORF 48, 78, or 100 can also inhibit such a homologous bacterial cellular component.

The demonstration that bacteriophage have adapted to inhibiting a host bacterium by acting on a particular cellular component or target provides a strong indication that that component is an appropriate target for developing and using antibacterial agents, e.g., in therapeutic treatments. Thus, the present invention provides additional guidance over mere identification of bacterial essential genes, as the present invention also provides an indication of accessibility of the target to an inhibitor, and an indication that the target is sufficiently stable over time (e.g., not subject to high rates of mutation) as phage acting on that target were able to develop and persist. Thus, the present invention identifies a particular subset of essential cellular components which are particularly likely to be appropriate targets for development of antibacterial agents.

The invention also, therefore, concerns the development or identification of inhibitors of bacteria, in addition to the phage-encoded inhibitory proteins (or RNA transcripts), which are active on the targets of bacteriophage-encoded inhibitors. As described herein, such inhibitors can be of a variety of different types, but are preferably small molecules.

The following description provides preferred methods for implementing the various aspects of the invention. However, as those skilled in the art will readily recognize, other approaches can be used to obtain and process relevant information. Thus, the invention is not limited to the specifically described methods. In addition, the following description provides a set of steps in a particular order. That series of steps describes the overall development involved in the present invention. However, it is clear that individual steps or portions of steps may be usefully practiced separately, and, further, that certain steps may be performed in a different order or even bypassed if appropriate information is already available or is provided by other sources or methods.

Identification of Inhibitory ORF

The methodology previously described in U.S. application Ser. No. 09/407,804 filed Sep. 28, 1999, and PCT International Application No. PCT/IB99/02040, was used to identify and characterize DNA sequences from Staphylococcus aureus bacteriophages 3A, 77 and 96 that can act as anti-microbials.

A nucleic acid segment isolated from Staphylococcus aureus bacteriophages 3A, 77 or 96 encodes a protein, whose gene is referred to as ORF (open reading frame) 33, 41, 79, 1, 48, 78, or 100 Thus, the present invention provides a nucleic acid sequence isolated from Staphylococcus aureus (Staph A or S. aureus) bacteriophages 3A, 77, or 96 comprising at least a portion of the gene encoding phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 with anti-microbial activity. The nucleic acid sequence can be isolated using a method similar to those described herein, or using another method. In addition, such a nucleic acid sequence can be chemically synthesized. Having the anti-microbial nucleic acid sequence of the present invention, parts thereof or oligonucleotides derived therefrom, other anti-microbial sequences from other bacteriophage sources using methods described herein or other methods can be isolated, including screening methods based on nucleic acid sequence hybridization.

The present invention provides the use of the Staph A bacteriophages 3A, 77, or 96 anti-microbial DNA segment encoding phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100, as a pharmacological agent, either wholly or in part, as well as the use of peptidomimetics, developed from amino acid or nucleotide sequence knowledge of Staph A phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. This can be achieved where the structure of the peptidomimetic compound corresponds to the structure of the active portion of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. In this analysis, the peptide backbone is transformed into a carbon-based hydrophobic structure that can retain cytostatic or cytocidal activity for the bacterium. This is done by standard medicinal chemistry methods, measuring growth inhibition of the various molecules in liquid cultures or on solid medium. These mimetics also represent lead compounds for the development of novel antibiotics.

In this context, “corresponds” means that the peptidomimetic compound structure has sufficient similarities to the structure of the active portion of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 that the peptidomimetic will interact with the same molecule as the product of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 and preferably will elicit at least one cellular response in common which relates to the inhibition of the cell by the phage protein.

The invention also provides bacteriophage anti-microbial DNA segments from other phages based on nucleic acids and sequences hybridizing to the presently identified inhibitory ORF under high stringency conditions or sequences which are homologous as described above. The bacteriophage anti-microbial DNA segment from phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 can be used to identify a related segment from another related or unrelated phage based on conditions of hybridization or sequence comparison.

Identification of Bacterial Targets

The present invention provides the use of Staphylococcus phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 anti-microbial activity to identify essential host bacterium interacting proteins or other targets that could, in turn, be used for drug design and/or screening of test compounds. Thus, the invention provides a method of screening for antibacterial agents by determining whether test compounds interact with (e.g., bind to) the bacterial target. The invention also provides a method of making an antibacterial agent based on production and purification of the protein or RNA product of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. The method involves identifying a bacterial target of the product of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100, screening a plurality of compounds to identify a compound active on the target, and synthesizing the compound in an amount sufficient to provide a therapeutic effect when administered to an organism infected by a bacterium naturally producing the target. The rationale is that the product of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 can physically interact and/or modify certain microbial host components to block their function.

A variety of methods are known to those skilled in the art for identifying interacting molecules and for identifying target cellular components. Several approaches and techniques are described below which can be used to identify the host bacterial pathway and protein that interact or are inhibited by phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100.

The first approach is a genetic screen for protein:protein interaction, e.g., either some form of two hybrid screen or some form of suppressor screen. In one form of the two hybrid screen involving the yeast two hybrid system, the nucleic acid segment encoding phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100, or a portion thereof, is fused to the carboxyl terminus of the yeast Gal4 DNA binding domain to create a bait vector. A genomic DNA library of cloned S. aureus sequences which have been engineered into a plasmid where the S. aureus sequences are fused to the carboxyl terminus of the yeast of Gal4 activation domain II (amino acids 768-881), is also generated. These plasmids are introduced alone, or in combination, into a yeast strain, e.g., Y190, previously engineered with chromosomally integrated copies of the E. coli lacZ and the selectable His3 genes, both under Gal4 regulation (Durfee et al., 1993). If the two proteins expressed in yeast interact, the resulting complex will activate transcription from promoters containing Gal4 binding sites. A lacZ and His3 gene, each driven by a promoter containing Gal4 binding sites, have been integrated into the genome of the host yeast system and are used for measuring protein-protein interactions. Such a system provides a physiological environment in which to detect potential protein interactions.

This system has been extensively used to identify novel protein-protein interaction partners and to map the sites required for interaction (for example, to identify interacting partners of translation factors (Qui et al., 1998), transcription factors (Katagiri et al., 1998), proteins involved in signal transduction (Endo et al., 1997). Alternatively, a bacterial two-hybrid screen can be utilized to circumvent the need for the interacting proteins to be targeted to the nucleus, as is the case in the yeast system (Karimova et al., 1998).

The protein targets of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 can also be identified using bacterial genetic screens. One approach involves the overexpression of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 protein in mutagenized S. aureus followed by plating the cells and searching for colonies that can survive the anti-microbial activity of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. These colonies are then grown, their DNA extracted, and cloned into an expression vector that contains a replicon of a different incompatibility group from the plasmid expressing phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. This library is then introduced into a wild-type Staph A bacterium in conjunction with an expression vector driving synthesis of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100, followed by selection for surviving bacteria. Thus, Staph A DNA fragments from the survivors presumably contain a DNA fragment from the original mutagenized Staph A genome that can protect the cell from the antimicrobial activity phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. This fragment can be sequenced and compared with that of the bacterial host to determine in which gene the mutation lies. This approach enables one to determine the targets and pathways that are affected by the killing function.

Alternatively, the bacterial targets can be determined in the absence of selecting for mutations using the approach known as “multicopy suppression”. In this approach, the DNA from the wild type Staph A host is cloned into an expression vector that can coexist with the one containing phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100. Those plasmids that contain host DNA fragments and genes which protect the host from the anti microbial activity of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 can then be isolated and sequenced to identify putative targets and pathways in the host bacteria.

Another approach is based on identifying protein:protein interactions between the product of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 and S. aureus host proteins, using a biochemical approach based on affinity chromatography. This approach has been used to identify interactions between lambda phage proteins and proteins from their E. coli host (Sopta et al., 1995). The product of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 is fused to a tag (e.g. -glutathione-S-transferase) after insertion in a commercially available plasmid vector which directs high-level expression after induction of the responsive promoter driving the fusion protein. The fusion protein is expressed in E. coli, purified, and immobilized on a solid phase matrix. Total cell extracts from S. aureus are then passed through the affinity matrix containing the immobilized phage ORF fusion protein; host proteins retained on the column are then eluted under different conditions of ionic strength, pH, and detergents and identified by gel electrophoresis. They are recovered from the gel by transfer to a high affinity membrane. The proteins are individually digested to completion with a protease (e.g.-trypsin) and either molecular mass or the amino acid sequence of the tryptic fragments can be determined by mass spectrometry using MALDI-TOF technology (Qin et al., 1997). The sequence of the individual peptides from a single protein are then analyzed by a bioinformatics approach to identify the S. aureus protein interacting with the phage ORF. This is performed by a computer search of the S. aureus genome for the identified sequence. Alternatively, tryptic peptide fragments of the S. aureus genome can be predicted by computer software based on the nucleotide sequence of the genome, and the predicted molecular mass of peptide fragments generated in silico compared to the molecular mass of the peptides obtained from each interacting protein eluted from the affinity matrix.

In addition, an oligonucleotide cocktail can be synthesized based on the primary amino acid sequence determined for an interacting S. aureus protein fragment. This oligonucleotide cocktail would comprise a mixture of oligonucleotides based on the nucleotide sequences of the primary amino acid of the predicted peptide, but in which all possible codons for a particular amino acid sequence are present in a subset of the oligonucleotide pool. This cocktail can then be used as a degenerate probe set to screen, by hybridization to genomic or cDNA libraries, to isolate the corresponding gene.

Alternatively, antibodies raised to peptides which correspond to an interacting S. aureus protein fragment can be used to screen expression libraries (genomic or cDNA) to identify the gene encoding the interacting protein.

Vectors

The invention also provides vectors, preferably expression vectors, harboring the anti-microbial DNA nucleic acid segment of the invention in an expressible form, and cells transformed with the same. Such cells can serve a variety of purposes, such as in vitro models for the function of the anti-microbial nucleic acid segment and screening for downstream targets of the anti-microbial nucleic acid segment, as well as expression to provide relatively large quantities of the inhibitory product.

Thus, an expression vector harboring the anti-microbial nucleic acid segment or parts thereof (Staph A bacteriophage 3A ORF 33, 41, 79, bacteriophage 77 ORF 1, bacteriophage 96 ORF 48, 78, 100) can also be used to obtain substantially pure protein. Well-known vectors, such as the pGEX series (available from Pharmacia), can be used to obtain large amounts of the protein which can then be purified by standard biochemical methods based on charge, molecular mass, solubility, or affinity selection of the protein by using gene fusion techniques (such as GST fusion, which permits the purification of the protein of interest on a glutathione column). Other types of purification methods or fusion proteins could also be used as recognized by those skilled in the art.

Likewise, vectors containing phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 can be used in methods for identifying targets of the encoded antibacterial ORF product, e.g., as described above, and/or for testing inhibition of homologous bacterial targets or other potential targets in bacterial species other than Staphylococcus aureus.

Antibodies

Antibodies, both polyclonal and monoclonal, can be prepared against the protein encoded by a bacteriophage anti-microbial DNA segment of the invention (e.g., Staph A phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100) by methods well known in the art. Protein for preparation of such antibodies can be prepared by purification, usually from a recombinant cell expressing the specified ORF or fragment thereof. Those skilled in the art are familiar with methods for preparing polyclonal or monoclonal antibodies (See, e.g., Antibodies: A Laboratory Manual, Harlow and Lane, Cold Spring Harbor Laboratory, CSHL Press, N.Y., 1988).

Such antibodies can be used for a variety of purposes including affinity purification of the protein encoded by the bacteriophage anti-microbial DNA segment, tethering of the protein encoded by the bacteriophage anti-microbial DNA segment to a solid matrix for purposes of identifying interacting host bacterium proteins, and for monitoring of expression of the protein encoded by the bacteriophage anti-microbial DNA segment.

Recombinant Cells

Bacterial cells containing an inducible vector regulating expression of the bacteriophage anti-microbial DNA segment can be used to generate an animal model system for the study of infection by the host bacterium. The functional activity of the proteins encoded by the bacteriophage anti-microbial DNA segments, whether native or mutated, can be tested in animal in vitro or in vivo models.

While such cells containing inducible expression vectors is preferred, other recombinant cells containing a recombinant phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 sequence or portion thereof are also provided by the present invention.

Also, a recombinant cell may contain a recombinant sequence encoding at least a portion of a protein which is a target of phage 3A ORF 33, 41, or 79, phage 77 ORF 1, or phage 96 ORF 48, 78, or 100 inhibitory ORF product.

In the context of this invention, in connection with nucleic acid sequences, the term “recombinant” refers to nucleic acid sequences which have been placed in a genetic location by intervention using molecular biology techniques, and does not include the relocation of phage sequences during or as a result of phage infection of a bacterium or normal genetic exchange processes such as bacterial conjugation.

Derivatization of Identified Anti-microbials

In cases where the identified anti-microbials above are peptidic compounds, the in vivo effectiveness of such compounds may be advantageously enhanced by chemical modification using the natural polypeptide as a starting point and incorporating changes that provide advantages for use, for example, increased stability to proteolytic degradation, reduced antigenicity, improved tissue penetration, and/or improved delivery characteristics.

In addition to active modifications and derivative creations, it can also be useful to provide inactive modifications or derivatives for use as negative controls or introduction of immunologic tolerance. For example, a biologically inactive derivative which has essentially the same epitopes as the corresponding natural antimicrobial can be used to induce immunological tolerance in a patient being treated. The induction of tolerance can then allow uninterrupted treatment with the active anti-microbial to continue for a significantly longer period of time.

Modified anti-microbial polypeptides and derivatives can be produced using a number of different types of modifications to the amino acid chain. Many such methods are known to those skilled in the art. The changes can include, for example, reduction of the size of the molecule, and/or the modification of the amino acid sequence of the molecule. In addition, a variety of different chemical modifications of the naturally occurring polypeptide can be used, either with or without modifications to the amino acid sequence or size of the molecule. Such chemical modifications can, for example, include the incorporation of modified or non-natural amino acids or non-amino acid moieties during synthesis of the peptide chain, or the post-synthesis modification of incorporated chain moieties.

The oligopeptides of this invention can be synthesized chemically or through an appropriate gene expression system. Synthetic peptides can include both naturally occurring amino acids and laboratory synthesized, modified amino acids.

Also provided herein are functional derivatives of anti-microbial proteins or polypeptides. By “functional derivative” is meant a “chemical derivative,” “fragment,” “variant,” “chimera,” or “hybrid” of the polypeptide or protein, which terms are defined below. A functional derivative retains at least a portion of the function of the protein, for example, reactivity with a specific antibody, enzymatic activity or binding activity.

A “chemical derivative” of the complex contains additional chemical moieties not normally a part of the protein or peptide. Such moieties may improve the molecule's solubility, absorption, biological half-life, and the like. The moieties may alternatively decrease the toxicity of the molecule, eliminate or attenuate any undesirable side effect of the molecule, and the like. Moieties capable of mediating such effects are disclosed in Genaro, 1995, Remington's Pharmaceutical Science. Procedures for coupling such moieties to a molecule are well known in the art. Covalent modifications of the protein or peptides are included within the scope of this invention. Such modifications may be introduced into the molecule by reacting targeted amino acid residues of the peptide with an organic derivatizing agent that is capable of reacting with selected side chains or terminal residues, as described below.

Cysteinyl residues most commonly are reacted with alpha-haloacetates (and corresponding amines), such as chloroacetic acid or chloroacetamide, to give carboxymethyl or carboxyamidomethyl derivatives. Cysteinyl residues also are derivatized by reaction with bromotrifluoroacetone, chloroacetyl phosphate, N-alkylmaleimides, 3-nitro-2-pyridyl disulfide, methyl 2-pyridyl disulfide, p-chloro-mercuribenzoate, 2-chloromercuri-4-nitrophenol, or chloro-7-nitrobenzo-2-oxa-1,3-diazole.

Histidyl residues are derivatized by reaction with diethylprocarbonate at pH 5.5-7.0 because this agent is relatively specific for the histidyl side chain. Para-bromophenacyl bromide also is useful; the reaction is preferably performed in 0.1 M sodium cacodylate at pH 6.0.

Lysinyl and amino terminal residues are reacted with succinic or other carboxylic acid anhydrides. Derivatization with these agents has the effect of reversing the charge of the lysinyl residues. Other suitable reagents for derivatizing primary amine-containing residues include imidoesters such as methyl picolinimidate; pyridoxal phosphate; pyridoxal; chloroborohydride; trinitrobenzenesulfonic acid; O-methylisourea; 2,4 pentanedione; and transaminase-catalyzed reaction with glyoxylate.

Arginyl residues are modified by reaction with one or several conventional reagents, among them phenylglyoxal, 2,3-butanedione, 1,2-cyclohexanedione, and ninhydrin. Derivatization of arginine residues requires that the reaction be performed in alkaline conditions because of the high pK_(a) of the guanidine functional group. Furthermore, these reagents may react with the groups of lysine as well as the arginine alpha-amino group.

Tyrosyl residues are well-known targets of modification for introduction of spectral labels by reaction with aromatic diazonium compounds or tetranitromethane. Most commonly, N-acetylimidizol and tetranitromethane are used to form O-acetyl tyrosyl species and 3-nitro derivatives, respectively.

Carboxyl side groups (aspartyl or glutamyl) are selectively modified by reaction carbodiimide (R′—N—C—N—R′) such as 1-cyclohexyl-3-(2-morpholinyl(4-ethyl) carbodiumide or 1-ethyl-3-(4-azonia-4,4-dimethylpentyl) carbodiimide. Furthermore, aspartyl and glutamyl residues are converted to asparaginyl and glutaminyl residues by reaction with ammonium ions.

Glutaminyl and asparaginyl residues are frequently deamidated to the corresponding glutamyl and aspartyl residues. Alternatively, these residues are deamidated under mildly acidic conditions. Either form of these residues falls within the scope of this invention.

Derivatization with bifunctional agents is useful, for example, for cross-linking component peptides to each other or the complex to a water-insoluble support matrix or to other macromolecular carriers. Commonly used cross-linking agents include, for example, 1,1-bis (diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters, for example, esters with 4-azidosalicylic acid, homobifunctional imidoesters, including disuccinimidyl esters such as 3,3′-dithiobis(succinimidylpropionate), and bifunctional maleimides such as bis-N-maleimido-1,8-octane. Derivatizing agents such as methyl-3-[p-azidophenyl) dithiolpropioimidate yield photoactivatable intermediates that are capable of forming crosslinks in the presence of light. Alternatively, reactive water-insoluble matrices such as cyanogen bromide-activated carbohydrates and the reactive substrates described in U.S. Pat. Nos. 3,969,287; 3,691,016; 4,195,128; 4,247,642; 4,229,537; and 4,330,440 are employed for protein immobilization.

Other modifications include hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of seryl or threonyl residues, methylation of the alpha-amino groups of lysine, arginine, and histidine side chains (Creighton, T. E., Proteins: Structure and Molecular Properties, W. H. Freeman & Co., San Francisco, pp. 79-86 (1983)), acetylation of the N-terminal amine, and, in some instances, amidation of the C-terminal carboxyl groups.

Such derivatized moieties may improve the stability, solubility, absorption, biological half-life, and the like. The moieties may alternatively eliminate or attenuate any undesirable side effect of the protein complex. Moieties capable of mediating such effects are disclosed, for example, in Genaro, 1995, Remington's Pharmaceutical Science.

The term “fragment” is used to indicate a polypeptide derived from the amino acid sequence of the protein or polypeptide having a length less than the full-length polypeptide from which it has been derived. Such a fragment may, for example, be produced by proteolytic cleavage of the full-length protein. Preferably, the fragment is obtained recombinantly by appropriately modifying the DNA sequence encoding the proteins to delete one or more amino acids at one or more sites of the C-terminus, N-terminus, and/or within the native sequence.

Another functional derivative intended to be within the scope of the present invention is a “variant” polypeptide which either lacks one or more amino acids or contains additional or substituted amino acids relative to the native polypeptide. The variant may be derived from a naturally occurring polypeptide by appropriately modifying the protein DNA coding sequence to add, remove, and/or to modify codons for one or more amino acids at one or more sites of the C-terminus, N-terminus, and/or within the native sequence.

A functional derivative of a protein or polypeptide with deleted, inserted and/or substituted amino acid residues may be prepared using standard techniques well-known to those of ordinary skill in the art. For example, the modified components of the functional derivatives may be produced using site-directed mutagenesis techniques (as exemplified by Adelman et al., 1983, DNA 2:183; Sambrook et al., 1989) wherein nucleotides in the DNA coding sequence are modified such that a modified coding sequence is produced, and thereafter expressing this recombinant DNA in a prokaryotic or eukaryotic host cell, using techniques such as those described above. Alternatively, components of functional derivatives of complexes with amino acid deletions, insertions and/or substitutions may be conveniently prepared by direct chemical synthesis, using methods well-known in the art.

Insofar as other anti-microbial inhibitor compounds identified by the invention described herein may not be peptidal in nature, other chemical techniques exist to allow their suitable modification, as well, and according the desirable principles discussed above.

Administration and Pharmnaceutical Compositions

For the therapeutic and prophylactic treatment of infection, the preferred method of preparation or administration of anti-microbial compounds will generally vary depending on the precise identity and nature of the anti-microbial being delivered. Thus, those skilled in the art will understand that administration methods known in the art will also be appropriate for the compounds of this invention. Pharmaceutical compositions are prepared, as understood by those skilled in the art, to be appropriate for therapeutic use. Thus, generally the components and composition are prepared to be sterile and free of components or contaminants which would pose an unacceptable risk to a patient. For compositions to be administered internally is is generally important that the composition be pyrogen free, for example.

The particularly desired anti-microbial can be administered to a patient either by itself, or in pharmaceutical compositions where it is mixed with suitable carriers or excipient(s). In treating an infection, a therapeutically effective amount of an agent or agents is administered. A therapeutically effective dose refers to that amount of the compound that results in amelioration of one or more symptoms of bacterial infection and/or a prolongation of patient survival or patient comfort.

Toxicity,. therapeutic and prophylactic efficacy of anti-microbials can be determined by standard pharmaceutical procedures in cell cultures and/or experimental organisms such as animals, e.g., for determining the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀. Compounds which exhibit large therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED₅₀ with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.

For any compound identified and used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. Such information can be used to more accurately determine useful doses in organisms such as plants and animals, preferably mammals, and most preferably humans. Levels in plasma may be measured, for example, by HPLC or other means appropriate for detection of the particular compound.

The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition (see e.g. Fingl et. al., in The Pharmacological Basis of Therapeutics, 1975, Ch. 1 p.1).

It should be noted that the attending physician would know how and when to terminate, interrupt, or adjust administration due to toxicity, organ dysfunction, or other systemic malady. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity). The magnitude of an administered dose in the management of the disorder of interest will vary with the severity of the condition to be treated and the route of administration. The severity of the condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency, will also vary according to the age, body weight, and response of the individual patient. A program comparable to that discussed above also may be used in veterinary or phyto medicine.

Depending on the specific infection target being treated and the method selected, such agents may be formulated and administered systemically or locally, i.e., topically. Techniques for formulation and administration may be found in Genaro, 1995, Remington's Pharmaceutical Science. Suitable routes may include, for example, oral, rectal, transdermal, vaginal, transmucosal, intestinal, parenteral, intramuscular, subcutaneous, or intramedullary injections, as well as intrathecal, intravenous, or intraperitoneal injections.

For injection, the agents of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

Use of pharmaceutically acceptable carriers to formulate identified anti-microbials of the present invention into dosages suitable for systemic administration is within the scope of the invention. With proper choice of carrier and suitable manufacturing practice, the compositions of the present invention, in particular those formulated as solutions, may be administered parenterally, such as by intravenous injection. Appropriate compounds can be formulated readily using pharmaceutically acceptable carriers well known in the art into dosages suitable for oral administration. Such carriers enable the compounds of the invention to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated.

Agents intended to be administered intracellularly may be administered using techniques well known to those of ordinary skill in the art. For example, such agents may be encapsulated into liposomes, then administered as described above. Liposomes are spherical lipid bilayers with aqueous interiors. All molecules present in an aqueous solution at the time of liposome formation are incorporated into the aqueous interior. The liposomal contents are both protected from the external microenvironment and, because liposomes fuse with cell membranes, are efficiently delivered into the cell cytoplasm. Additionally, due to their hydrophobicity, small organic molecules may be directly administered intracellularly.

Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art.

In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. The preparations formulated for oral administration may be in the form of tablets, dragees, capsules, or solutions, including those formulated for delayed release or only to be released when the pharmaceutical reaches the small or large intestine.

The pharmaceutical compositions of the present invention may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levitating, emulsifying, encapsulating, entrapping or lyophilizing processes.

Pharmaceutical formulations for parenteral administration include aqueous solutions of the active anti-microbial compounds in water-soluble form. Alternatively, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

Pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added.

The above methodologies may be employed either actively or prophylactically against an infection of interest.

To identify DNA segments of Staph A bacteriophages 3A, 77 and 96 capable of acting as anti-microbial agents, a strategy described in U.S. application Ser. No. 09/407,804 filed Sep. 28, 1999, and PCT International Application No. PCT/IB99/02040 was employed. In essence, the procedure involved sequence characterization of the bacteriophage, identification of protein coding regions (open reading frames or ORFs), subcloning of all ORFs into an appropriate inducible expression vector, transfer of the ORF subclones into Staph. A, followed by induction of ORF expression and assessment of effect on growth. We employed discovery steps as described in the Examples.

EXAMPLE I Growth of Staphylococcus aureus Bacteriophages 3A, 77 and 96 and Purification of Genomic DNA

The Staphylococcus aureus propagating strain (PS 3A, 77 and 96) (Laboratory Center for Disease Control. (CDC) Health Canada, Ottawa, Ontario) were used as host to propagate their respective phages 3A, 77 and 96, also obtained from the CDC. Two rounds of plaque purification of phages were performed on soft agar essentially as described in Sambrook et al (1989). Briefly, the PS 3A, 77 and 96 strains were grown overnight at 37° C. in Nutrient broth [NB: 0.3% Bacto beef extract, 0.5% Bacto peptone (Difco Laboratories) and 0.5% NaCl (w/v)]. The culture was then diluted 20× in NB and incubated at 37° C. until the OD₅₄₀=0.2 (early log phase) with constant agitation. In order to obtain single plaques, phages 3A, 77 or 96 were subjected to 10-fold serial dilutions using phage buffer (1 mM MgSO₄, 5 mM MgCl₂, 80 mM NaCl and 0.1% Gelatin (w/v)) and 10 μl of each dilution was used to infect 0.5 ml of the cell suspension in the presence of 400 μg/ml CaCl₂. After incubation of 15 min at room temperature (RT), 2 ml of melted soft agar kept at 45° C. (NB supplemented with 0.6% agar) was added to the mixture and poured onto the surface of 100 mm nutrient agar plates (0.3% Bacto Beef extract, 0.5% Bacto peptone, 0.5% NaCl and 1.5% Bacto agar (w/v)). After overnight incubation at 30° C., a single plaque was isolated, resuspended in 1 ml of phage buffer by end over end rotation for 2 hrs at 20° C., and the phage suspension was diluted and used for a second infection as described above. After overnight incubation at 30° C., a single plaque was isolated and used as a stock.

The propagation procedure for bacteriophages 3A, 77 and 96 was modified from the agar layer method of Swanstörm and Adams (1951). Briefly, the respective PS strains were grown to stationary phase overnight at 37° C. in Nutrient broth. Each culture was then diluted twenty-fold in NB and incubated at 37° C. until the OD₅₄₀=0.2. The suspension (15×10⁷ Bacteria) was then mixed with 15×10⁵ plaque forming units (pfu) to give a ratio of 100-bacteria/phage particle in the presence of 400 μg/ml of CaCl₂. After incubation for 15 min at 20° C., 7.5 ml of melted soft agar (NB plus 0.6% agar) were added to the mixture and poured onto the surface of 150 mm nutrient agar plates and incubated 16 hrs at 37° C. To collect the phage plate lysate, 20 ml of NB were added to each plate and the soft agar layer was collected by scrapping off with a clean microscope slide followed by shaking of the agar suspension for 5 min to break up the agar. The mixture was then centrifuged for 10 min at 4,000 RPM (2,830×g) in a JA-10 rotor (Beckman) and the supernatant fluid (lysate) was collected and subjected to a treatment with 10 μg /ml of DNase I and RNase A for 30 min at 37° C. To precipitate the phage particles, the phage suspension was adjusted to 10% (w/v) PEG 8000 and 0.5 M of NaCl followed by incubation at 4° C. for 16 hrs. The phage was recovered by centrifugation at 4,000 rpm (3,500×g) for 20 min at 4° C. on a GS-6R table top centrifuge (Beckman). The pellet was resuspended with 2 ml of phage buffer (1 mM MgSO₄, 5 mM MgCl₂, 80 mM NaCl and 0.1% Gelatin). The phage suspension was extracted with 1 volume of chloroform and further purified by centrifugation on a cesium chloride step gradient as described in Sambrook et al. (1989), using a TLS 55 rotor centrifuged in an Optima TLX ultracentrifuge (Beckman) for 2 h at 28,000 rpm (67,000×g) at 4° C. Banded phage was collected and ultracentrifuged again on an isopycnic cesium chloride gradient (1.45 g/ml) at 40,000 rpm (64,000×g) for 24 h at 4° C. using a TLV rotor (Beckman). The phage was harvested and dialyzed for 4 h at room temperature against 4 L of dialysis buffer consisting of 10 mM NaCl, 50 mM Tris-HCl [pH 8] and 10 mM MgCl₂. Phage DNA was prepared from the phage suspension by adding 20 mM EDTA, 50 ug/ml Proteinase K and 0.5% SDS and incubating for 1 h at 65° C., followed by successive extractions with 1 volume of phenol, 1 volume of phenol-chloroform and 1 volume of chloroform. The DNA was then dialyzed overnight at 4° C. against 4 L of TE (10 mM Tris-HCl [pH 8.0], 1 mM EDTA).

EXAMPLE II DNA Sequencing of Bacteriophage 3A, 77 and 96 Genomes

Four micrograms of phage DNA was diluted in 200 μl of TE, 1 mM EDTA in a 1.5 ml eppendorf tube and sonication was performed (550 Sonic Dismembrator™, Fisher Scientific). Samples were sonicated under an amplitude of 3 μm with bursts of 5 s spaced by 15 s cooling in ice/water for 3 to 4 cycles. The sonicated DNA was then size fractionated by electrophoresis on 1% agarose gels utilizing TAE (1×TAE is: 40 mM Tris-acetate, 1 mM EDTA [pH 8.0]) as the running buffer. Fractions ranging from 1 to 2 kbp were excised from the agarose gel and purified using a commercial DNA extraction system according to the instructions of the manufacturer (Qiagen), with a final elution of 50 μl of 1 mM Tris-HCl [pH 8.5].

The ends of the sonicated DNA fragments were repaired with a combination of T4 DNA polymerase and the Klenow fragment of E. coli DNA polymerase 1, as follows. Reactions were performed in a reaction mixture (final volume, 100 μl) containing sonicated phage DNA, 10 mM Tris-HCl [pH 8.0], 50 mM NaCl, 10 mM MgCl₂, 1 mM DTT, 50 μg/ml BSA, 100 μM of each dNTP and 15 units of T4 DNA polymerase (New England Biolabs) for 20 min at 12° C. followed by addition of 12.5 units of Klenow large fragment (New England Biolabs) for 15 min at room temperature. The reaction was stopped by two phenol/chloroform extractions and the DNA was precipitated with ethanol and the final DNA pellet was resuspended in 20 μl of H₂O.

Blunt-ended DNA fragments were cloned by ligation directly into the Hinc II site of pKSII+ vector (Stratagene) dephosphorylated by treatment with calf intestinal alkaline phosphatase (New England Biolabs). A typical ligation reaction contained 100 ng of vector DNA, 2 to 5 μl of repaired sonicated phage DNA (50-100 ng) in a final volume of 20 μl containing 800 units of T4 DNA ligase (New England Biolabs) and was incubated overnight at 16° C. Transformation and selection of bacterial clones containing recombinant plasmids was performed in E. coli DH10β according to standard procedures (Sambrook et al., 1989).

Recombinant clones were picked from agar plates into 96-well plates containing 100 μl LB and 100 μg/ml ampicillin and incubated at 37° C. The presence of phage DNA insert was confirmed by PCR amplification using T3 and T7 primers flanking the Hinc II cloning site of the pKS II+ vector. PCR amplification of foreign insert was performed in a 15 μl reaction volume containing 10 mM Tris-HCl [pH 8.3], 50 mM KCl, 1.5 mM MgCl₂, 0.02% gelatin, 1 μM primer, 187.5 μM each dNTP, and 0.75 units Taq polymerase (BRL). The thermocycling parameters were as follows: 2 min initial denaturation at 94° C. for 2 min, followed by 20 cycles of 30 sec denaturation at 94° C., 30 sec annealing at 57° C., and 2 min extension at 72° C., followed by a single extension step at 72° C. for 10 min. Clones with insert sizes of 1 to 2 kbp were selected and plasmid DNA was prepared from the selected clones using QIAprep™ spin miniprep kit (Qiagen).

The nucleotide sequence of the extremities of each recombinant clone was determined using an ABI 377-36 automated sequencer with two types of chemistry:ABI prism Big Dye™ primer cycle sequencing (21M13 primer: #403055)(M13REV primer: #403056) or ABI prism Big Dye™ terminator cycle sequencing ready reaction kit (Applied Biosystems, #4303152). To ensure co-linearity of the sequence data and the genome, all regions of phage genome were sequenced at least once from both directions on two separate clones. In areas that this criteria was not initially met, a sequencing primer was selected and phage DNA was used directly as sequencing template employing ABI prism Big Dye™ terminator cycle sequencing ready reaction kit.

EXAMPLE III Bioinformatic Management of Primary Nucleotide Sequence

Sequence contigs were assembled using Sequencher™ 3.1 software (GeneCodes). To close contig gaps, sequencing primers were selected near the edge of the contigs. Phage DNA was used directly as sequencing template employing ABI prism BIG DYE™ terminator cycle sequencing ready reaction kit. The complete sequences of bacteriophages 3A, 77 and 96 are shown in Table 1.

A software program was developed and used on the assembled sequence of the bacteriophages to identify all putative ORFs larger than 33 codons. Other ORF identification software can also be utilized, preferably programs which allow alternative start codons. The software scans the primary nucleotide sequence starting at nucleotide #1 for an appropriate start codon. Three possible selections can be made for defining the nature of the start codon; I) selection of ATG, II) selection of ATG or GTG, and III) selection of either ATG, GTG, TTG, CTG, ATT, ATC, and ATA. This latter initiation codon set corresponds to the one reported by the NCBI (http://www.ncbi.nlm.nih.gov/htbin-post/Taxonomy/wprintyc?mode=c) for the bacterial genetic code.

When an appropriate start codon is encountered, a counting mechanism is employed to count the number of codons (groups of three nucleotides) between this start codon and the next stop codon downstream of it. If a threshold value of 33 is reached, or exceeded, then the sequence encompassed by these two codons (start and stop codons) is defined as an ORF. This procedure is repeated, each time starting at the next nucleotide following the previous stop codon found, in order to identify all the other putative ORFs. The scan is performed on all three reading frames of both DNA strands of the phage sequence.

Sequence homology (BLAST) searches for each ORF are then carried out using an implementation of BLAST programs, although any of a variety of different sequence comparison and matching programs can be utilized as known to those skilled in the art. Downloaded public databases used for sequence analysis include:

i) non-redundant GenBank (ftp://ncbi.nlm.nih.gov/blast/db/nr.Z),

ii) Swissprot (ftp://ncbi.nlm.nih.gov/blast/db/swissprot.Z);

iii) vector (ftp://ncbi.nlm.nih.gov/blast/db/vector.Z);

iv) pdbaa databases (ftp://ncbi.nlm.nih.gov/blast/db/pdbaa.Z);

v) staphylococcus aureus NCTC 8325 (ftp://ftp.genome.ou.edu/pub/staph/staph-1k.fa);

vi) streptococcus pyogenes (ftp://ftp.genome.ou.edu/pub/strep/strep-1k.fa);

vii) streptococcus pneumoniae

(ftp://ftp.tigr.org/pub/data/s_neumoniae/gsp.contigs. 112197.Z);

viii) mycobacterium tuberculosis CSU#9

(ftp://ftp.tigr.org/pub/data/m_tuberculosis/TB_(—)091097.Z) and ix)

pseudomonas aeruginosa

(http://www.genome.washington.edu/pseudo/data.html).

The results of the homology searches performed on the bacteriophage 3A, 77 and 96 ORFs are shown in Table 4.

EXAMPLE IV Subcloning of Bacteriophage 3A, 77 and 96 ORFs into a Staph A Inducible Expression System

Preparation of the Shuttle Expression Vectors

The shuttle vector pT0021, in which the firefly luciferase (lucFF) expression is controlled by the ars (arsenite) promoter/operator (Tauriainen et al., 1997), was modified as below to suit our specific application. Two oligonucleotides were synthesized. The sense strand sequence (with XhoI cloning site) is: 5′-AATTCTCGAGTAAAATAACAT-3′ (SEQ ID NO. 1); the antisense strand sequence (with a BamHI cloning site) is: 5′-CGGGATCCGCCTCCTTTTCTCAACAGTCACCTGATTT-3′ (SEQ ID NO. 2). The two oligonucleotides were used for polymerase chain reaction (PCR) amplification of pT0021 vector. The PCR product was gel purified using the Qiagen kit as described, and digested with XhoI and BamHI. The digested PCR product was again gel purified, ligated into XhoI and BamHI digested pT0021 vector, and used to transform E. coli bacterial strain DH10β (as described above). This manipulation results in the construction of a pT0021-intermediated vector containing a RBS sequence located immediately upstream of the BamHI cloning site. Two other oligonucleotides were synthesized. The sense strand sequence (with BamHI cloning site) is: 5′-CGGGATCCATGAGGGGTTCCGAAGACG-3′ (SEQ ID NO. 3); the antisense strand sequence (with a HindIII cloning site) is: 5′-CCCAAGCTTACAATTTGGACTTTC-3′ (SEQ ID NO. 4). The two oligonucleotides were used for PCR amplification of pT0021-intermediated vector. The PCR product was gel purified and digested with BamHI and HindIII. The digested PCR product was then gel purified as described, ligated into BamHI and HindIII digested pT0021-intermediated vector, and used to transform E. coli bacterial strain DH10β. This modified shuttle vector containing the ATG of the lucFF gene located immediately downstream of the BamHI cloning site was named pTM. A diagram outlining our modification of pT0021 to generate pTM is shown in FIG. 1A. The pTMSM vector is a modified version of the pTM vector containing the SalI and MluI cloning sites replacing the HindIII cloning site as shown in FIG. 1B. These modified shuttle vectors contain the arsenite inducible promoter/operator and the arsR gene.

As another example of inducible promotor, the arsenite-inducible promotor and the asrR gene from the pTMSM vector were replaced by a lactose-inducible promotor and the lacR gene from Staphylococcus aureus. The S. aureus gene encoding for the repressor of the lac operon (lacR) is found immediately upstream of the promoter-proximal end of the the lacA-G genes. Two oligonucleotides corresponding to a 2.18 kb-DNA region encompassing the lacR and the lac operon promotor region were synthesized. The sense strand sequence is: 5′-ccgctcgagCTCCAAATTCCAAAACAG-3′ (SEQ ID NO. 11) (with a XhoI cloning site, ctcgag); the antisense strand sequence is: 5′-cgggatccAATAAGACTCCTTTTTAC-3′ (SEQ ID NO. 12) (with a BamHI cloning site, ggatcc). These two oligonucleotides were used for the PCR amplification of Staphylococcus aureus DNA. The PCR product was gel purified and digested with XhoI and BamHI. The digested PCR product was also gel purified, ligated into XhoI and BamHI-digested pTMSM vector, and used to transform E. coli bacterial strain DH10β. In the resulting vector, pTMSLac, the firefly luciferase (lucFF) expression is under the control of the S. aureus lac operon promoter/operator. Recombinant pTMSLac clones were picked and the sequence integrity of the 2.1 8 kb-lac operon region (lacR+lac promotor) was verified directly by DNA sequencing. A diagram outlining the pTMSLac vector characteristics is shown in FIG. 1D.

For the analysis of the inhibitory ORFs expression in S. aureus, the pT0021 vectors was modified in the following fashion. Two oligonucleotides corresponding to a short antigenic peptide derived from the heamaglutinin protein of influenza virus (HA epitope tag) were synthesized (Field et al., 1988). The sense strand HA tag sequence (with BamHI, SalI and HindIII cloning sites) is: 5′-gatcccggtcgaccaagcttTACCCATACGACGTCCCAGACTACGCCAGCTGA-3′ (SEQ ID NO. 9) (where upper case letters denote the nucleotide sequence of the HA tag); the antisense strand HA tag sequence (with a HindIII cloning site) is: 5′-agctTCAGCTGGCGTAGTCTGGGACGTCGTATGGGTAaagcttggtcgaccgg-3′ (SEQ ID NO. 10) (where upper case letters denote the sequence of the HA tag). The two HA tag oligonucleotides were annealed and ligated into pT0021 vector which had been digested with BamHI and HindIII. This manipulation resulted in replacement of the lucFF gene by the HA tag. This modified shuttle vector containing the arsenite inducible promoter, the arsR gene, and HA tag was named pTHA. A diagram outlining our modification of pT0021 to generate pTHA is shown in FIG. 1C.

Cloning of ORFs With a Shine-Dalgarno Sequence.

Individual ORF, encoded by Bacteriophages 3A, 77 and 96, larger than 33 amino acids and having a Shine-Dalgarno sequence upstream of the initiation codon was selected for functional analysis. In total, 52 ORFs from phage 3A, 99 ORFs from phage 77 and 45 ORFs from phage 96 were selected and screened as detailed below. A list of these is presented in FIG. 4A. Each individual ORF, from initiation codon to stop codon was amplified from phage genomic DNA using the polymerase chain reaction (PCR). For PCR amplification of ORFs, each sense strand primer targets the initiation codon and is preceded by a BamHI restriction site (5′-cgggatcc-3′) and each antisense oligonucleotide targets the stop codon of the ORF and is preceded by a HindIII restriction site (5′-cccaagctt-3′) The PCR product of each ORF was purified using the Quiagen kit as described and digested with BamHI and HindIII. The digested PCR product was also purified using the Quiagen kit, ligated into BamHI and HindIII digested pTM vector and used to transform E. coli bacterial strain DH10β (as described above). As a result of this manipulation, the ORF is under the control of the arsenite-inducible promotor. Recombinant pTM/ORF clones were picked and their insert sizes were confirmed by PCR analysis using primers flanking the cloning site. The names and sequences of the primers that were used for the PCR amplification were: HAF: 5′-TATTATCCAAAACTTGAACA-3′ (SEQ ID NO. 14); HAR: 5′-CGGTGGTATATCCAGTGATT-3′ (SEQ ID NO. 15). The sequence integrity of cloned ORFs was verified directly by DNA sequencing using primers HAF and HAR. In cases where verification of ORF sequence could not be achieved by one pass with the sequencing primers, additional internal primers were selected and used for sequencing. In cases of ORF harboring internal HindIII site in their sequence, SalI instead of HindIII cloning site was used for the ORF cloning into the BamHI and SalI digested pTMSM vector. For the cloning into the lactose-inducible vector, the ORFs were excised from pTMSM vector by BamHI and SalI digestion and ligated to the same cloning sites into pTMSLac vector.

For the cloning into pTHA vector, each inhibitory ORF, from initiation codon to last codon (excluding the stop codon), was amplified from phage genomic DNA using the PCR. For PCR amplification of ORFs, each sense strand primer targets the initiation codon and is preceded by a BamHI restriction site (5′-cgggatcc-3′) and each antisense oligonucleotide targets the pentultimate codon (the one before the stop codon) of the ORF and is preceded by a Sal I restriction site (5′-gcgtcgaccg-3′) SEQ ID NO. 36). The PCR product of each ORF was gel purified and digested with BamHI and SalI. The digested PCR product was purified using the Qiagen kit as described, ligated into BaHI and SalI digested pTHA vector, and used to transform E. coli bacterial strain DH10β. As a result of this manipulation, the HA tag is set inframe with the ORF and is positioned at the carboxy terminus of each ORF (pTHA/ORF clones). Recombinant pTHA/ORF clones were picked and their insert sizes were confirmed as described above.

EXAMPLE V Functional Assay for Bacterial Inhibitory Activity of Bacteriophage 3A, 77 and 96 ORFs

Transformation of Staphylococcus aureus With Expression Construct Staphylococcus aureus strain RN4220 (Kreiswirth et al., 1983) was used as a recipient for the expression of recombinant plasmids. Electoporation was performed essentially as previously described (Schenk and Laddaga, 1992). Selection of recombinant clones was performed on Luria-Broth agar (LB-agar) plates containing 30 μg/ml of kanamycin.

For each ORF introduced in the pTM vector, 3 independent transformants were isolated and used to individually inoculate cultures in 5 ml of TSB containing 30 μg/ml kanamycin, followed by growth to saturation (16 hrs at 37° C.). An aliquot of this stationary phase culture was used to generate a frozen glycerol stock of the transformant (stored at −80° C.). With certain phage ORF, e.g. by phage 77 ORF 1 and 96 ORF 78, no S. aureus transformants could be obtained following cloning into pTM or pTMSM vector. In these cases, phage ORFs were cloned in alternative vectors pTHA and pTMSLac.

The presence of individual phage 3A, 77 or 96 ORF DNA inserts in the plasmid was verified by PCR amplification using 1.5 μl transformant miniprep DNA in a PCR with primers flanking the cloning site of ORF in pTM vector (HAF and HAR). The composition of the PCR reaction and the cycling parameters are identical to those employed for library screening described above.

Induction of Gene Expression From the ars- and lac-Inducible Promotors

Sodium arsenite (NaAsO₂) was purchased from Sigma (Sigma-Aldrich Canada LTD, Oakville) and was used as heavy metals to induce gene expression from the ars promoter/operator in solid and liquid medium assays.

The lactose (lac) genes of Staphylococcus aureus have been shown to be inducible with the addition of either lactose or galactose to the culture medium (Oskouian & Stewart, 1990, J. Bacteriol. 172 3804-3812). Galactose (2%w/v) was used to induce the gene expression from the lac promotor/operator in liquid assay.

At pre-determined times, sodium arsenite or galactose was added to the culture to induce transcription of the phage ORFs cloned immediately downstream from an arsenite-inducible promoter in the expression plasmids pTM, pTMSM or pTHA, or a lactose-inducible promotor in the expression plasmid pTMSLac. The anti-microbial activity of individual phage 3A, 77 and 96 ORFs was monitored by two growth inhibitory assays, one on solid agar medium, the other in liquid medium.

a-Screening on Semi-solid Support Media

ORFs were first screened by the functional assay on semi-solid medium as outlined in FIG. 3A. Cells containing different recombinant plasmids were grown overnight at 37° C. in LB medium supplemented with 30 μg/ml of kanamycin. The cells were then diluted and the identification of inhibitory ORFs was performed by spotting 3 ul of each dilution of S. aureus transformed cells containing phage 3A, 77 or 96 ORFs onto agar plates containing increasing concentrations of sodium arsenite (0; 2.5; 5; and 7.5 μM) and Kanamycin. The plates were incubated overnight at 37° C., after which a growth inhibition of the ORF transformants on plates that contain arsenite are compared to plates without arsenite. Noninduced and induced cultures of S aureus transformed with a non-inhibitory ORF (44AHJD bacteriophage ORF 114 cloned into pTM vector) were included as negative control. The 44AHJD ORF 114 amino acids residue composition from N-terminal to C-terminal is:MVNVDNAPEEKGQAYTEMLQLFNKLIQWNPAYTFDNAINLLSACQQLLLNYNSSVVQFLNDE LNNETKPESILSYIAGDDPIEQWNMHKGFYETYNVYVF (SEQ ID NO. 16).

Results of the bacteriophage ORFs tested for functional assay on semi-solid media are listed in FIG. 4A. Among them, induction of expression of phage 3A ORF 33, 41 or 79, phage 77 ORF 1, or phage 96 ORF 48 or 100 results in the inhibition of growth of the S. aureus transformants. FIG. 4B shows the result of growth inhibition with three clones of S. aureus expressing these inhibitory ORFs or the control non-inhibitory 44AHJD ORF 114.

b-Quantification of Growth inhibition in Liquid Medium

As outlined in FIG. 3B, the effect of ORF induction on bacterial growth inhibition was then further quantitated by functional assay in liquid medium. Cells containing phage 3A ORF 33, 41 or 79, phage 77 ORF 1, or phage 96 ORF 48, 78 or 100 were grown for overnight at 37° C. in LB medium supplemented with the appropriate antibiotic selection. These cultures were 50-fold dilution with fresh media containing kanamycin and the growth was continued for 2 h at 37° C. The same OD₅₆₅ equivalent of cultures (approximately 1 ml) was added to 19 ml of fresh media containing kanamycin and transferred to a 125 ml-Erlenmeyer flask. The cultures were incubated for an additional 4 hrs at 37° C. in the absence or in the presence of inducer (sodium arsenite at the final concentrations of 5.0 μM or 2.0% galactose). During that period of time, the effect of expression of the phage 3A, 77 and 96 ORFs on bacterial cell growth was monitored, at each 40 min, by measuring the OD₅₆₅ and the number of colony forming units (CFU) in the cultures containing or not the inducer. The number of CFU was evaluated as followed. Cultures were serially diluted and aliquots from induced and uninduced cultures were plated out on agar plates containing an appropriate antibiotic selection but lacking inducer. Following incubation overnight at 37° C., the number of colonies was counted. Cultures of S aureus transformed with a non-inhibitory ORF (44AHJD bacteriophage ORF 114 cloned into pTM vector) were included as control.

As shown in FIG. 5, for each inhibitory ORFs, the number of CFU increased over time under non-induced conditions. Similar growth rates were also observed with transformants harboring non-inhibitory ORFs under both induced and non-induced conditions. Transformants of S.aureus harboring C) phage 3A ORF 79 or D) phage 77 ORF 1 showed a significantly lower growth rate compared to their respective control cultures grown under non-induced conditions. Induction of expression of E) phage 96 ORF 100 was cytostatic. In contrast, four phage ORFs were cytocidal for bacterial growth. The expression of B) phage 3A ORF 41 resulted in a very rapid decrease in the number of viable cells as assayed as CFU. A 2 log reduction in the number of CFU after 1 hr of growth compared to the number of CFU initially present in the same culture was observed following induction of 3A ORF 41 with sodium arsenite. At 4 hr following induction, the number of viable cells relative to uninduced cultures was reduced by either 2 logs (phage 3A ORF 33 (A)), 1 log (phage 96 ORF 48 (D)), or 0.5 log (phage 96 ORF 78 (E)).

The presence of four phage ORFs were cytocydal for the bacterial growth. The expression of B) phage 3A ORF 41 results in a very rapid decrease in the number of CFU. A 2 log reduction in the number of CFU compared to the number of CFU initially present in the same culture was observed at 1 h following induction with sodium arsenite.

At 4 h following induction with sodium arsenite, the expression of A) phage 3A ORF 33 results in a 2 log reduction in the number of CFU compared to the number of CFU initially present in the same culture. The expression of D) phage 96 ORF 48 results in a log reduction in the number of CFU compared to the number of CFU initially present in the same culture.

At 4 h following induction of the expression of E) phage 96 ORF 78 with galactose a half log reduction in the number of CFU compared to the number of CFU initially present in the same culture was observed.

EXAMPLE VI Phage ORF Protein Expression Analysis in S. aureus

The level of expression of the inhibitory ORFs was measured by performing Western blot analyses. Staphylococcus aureus strain RN4220 was electroporated with each inhibitory ORFs cloned into pTHA vector as described above. Cells containing different recombinant plasmids were grown for overnight at 37° C. in TSB (Tryptic soy broth, DIFCO) medium in the presence of 30 μg/ml kanamycin. The overnight cultures were subjected to a 50-fold dilution with fresh media containing kanamycin and the growth was continued for 2 h at 37° C. At the end, cells were diluted with fresh TSB medium containing or not 5.0 μM of Sodium Arsenite, in the presence of kanamycin and incubated at 37° C. for an additional 3.5 h. The same OD₅₆₅ equivalent of cultures was centrifuged at 3000 g for 5 min and washed with 20 ml of TBS buffer (140 mM NaCl, 25 mM Tris-HCl, pH 7.5) containing protease inhibitors (1 mM of each phenylmethylsulfonyl fluoride (PMSF) and N-ethylmalemyde (NEM)). For lysis, cell pellets were resuspend in 25 μl with TBS buffer containing 1 mM PMSF, 1 mM NEM, 20 μg of each DNAse I and RNase A and 50 Units/ml of lysostaphin, and incubated at 37° C. for 1 h. The reaction was stopped by the addition of 25 μl of 2×SDS buffer (100 mM Tris pH 6.8, 4% SDS, 200 mM DTT, 20% Glycerol and 0.2% Bromophenol blue). Cell lysates were boiled for 10 min, centrifuged for 10 min at 13,000 g and 10-15 μl of the lysates were loaded onto a 15-18% SDS-page using Tris-Glycine-SDS as a running buffer (3.03 g of Tris HCl, 14.4 g of Glycine and 0.1% SDS per liter). After migration, proteins were transferred onto an immobilon-P membrane (PVDF, Millipore) using Tris-Glycin-Methanol as a transfer buffer (3.03 g Tris, 14.4 Glycine and 200 ml Methanol per liter) for 2 hrs at 4° C. at 100 V. PVDF membrane was pretreated in methanol for 30 s, washed 4-5 times with H₂O and soaked in transfer buffer.

After the transfer, the membrane was blocked in 20 ml of TBS containing 0.05% Tween-20 (TBST), 5% skim milk and 0.5% gelatin for 1 hr at room temperature and then, a pre-blocking antibody (ChromPureRabbit IgG, Jackson immunoResearch lab. #011-000-003) was added at a dilution of 1/750 and incubated for 1 hr at room temperature or ON at 4° C. Membrane was washed 6 times for 5 min in TBST at room temperature. The primary antibody (murine mono-HA antibody, Babco # MMS-101 P) directed against the HA epitope tag and diluted 1/1000 was then added and incubated for 3 h at room temperature in the presence of 5% Skim Milk and 0.5% Gelatin. Membrane was washed 6 times for 5 min in TBST at room temperature. A secondary antibody (anti-mouse IgG, peroxidase-linked species-specific whole antibody, Amersham # NA 931) diluted 1/1500 (7.5 μl in 10 ml) was then added and incubated for 1 hr at room temperature. After 6 washes in TBST, the membrane was briefly dried and then, the substrate (Chemiluminescence reagent plus, Mandel # NEL104) was added to the membrane and incubated for I min at room temperature. The membrane was briefly dried and exposed to x-ray film (Kodak, Biomax MS/MR ) for different periods of time (30 s to 10 min). As shows in FIG. 6, the presence of sodium arsenite in the cultures induces the expression of proteins corresponding to the phage 3A ORF 33, 41 and 79, phage 77 ORF 1, and phage 96 ORF 48, 78 and 100.

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All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually.

One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The specific methods and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. One of ordinary skill in the art would recognize that Bacteriophages 3A, 77 and 96 ORFs described herein are provided and discussed by way of example, and other the ORFs of Bacteriophages 3A, 77 and 96, including amino acid sequences and nucleic acid sequences which encode products, are within the scope of the present invention. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. For example, those skilled in the art will recognize that the invention may suitably be practiced using a variety of different expression vectors and sequencing methods within the general descriptions provided.

The invention illustratively described herein suitably may be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising,” “consisting essentially of” and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is not intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims.

In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group. For example, if there are alternatives A, B, and C, all of the following possibilities are included: A separately, B separately, C separately, A and B, A and C, B and C, and A and B and C.

Thus, additional embodiments are within the scope of the invention and within the following claims.

TABLE 2 1st 3rd position 2nd position position (5′ end) U C A G (3′ end) U Phe Ser Tyr Cys U Phe Ser Tyr Cys C Leu Ser Stop Stop A Leu Ser Stop Trp G C Leu Pro His Arg U Leu Pro His Arg C Leu Pro Gln Arg A Leu Pro Gln Arg G A Ile Thr Asn Ser U Ile Thr Asn Ser C Ile Thr Lys Arg A Met Thr Lys Arg G G Val Ala Asp Gly U Val Ala Asp Gly C Val Ala Glu Gly A Val Ala Glu Gly G

TABLE 3 3AORF033, Nucleotides and amino acids sequences (SEQ ID NO. 20) 30089 atggcaatattagaaggtatttttgaagaattaaaactattaaat 1 M  A  I  L  E  G  I  F  E  E  L  K  L  L  N 30134 aagaatttacgtgtgctaaatactgaactatcaactgtagattca 16 K  N  L  R  V  L  N  T  E  L  S  T  V  D  S 30179 tcaattgtacaagagaaagttaaagaagcaccaatgccaaaagat 31 S  I  V  Q  E  K  V  K  E  A  P  M  P  K  D 30224 gaaacagctcaactggaatcagttgaagaagttaaggaaacttct 46 E  T  A  Q  L  E  S  V  E  E  V  K  E  T  S 30269 gctgatttaactaaagattatgttttatcagtaggaaaagagttc 61 A  D  L  T  K  D  Y  V  L  S  V  G  K  E  F 30314 cttaaaaaagcagatacttctgataagaaagaatttagaaataaa 76 L  K  K  A  D  T  S  D  K  K  E  F  R  N  K 30359 cttaacgaacttggtgcggataagctatctactatcaaagaagag 91 L  N  E  L  G  A  D  K  L  S  T  I  K  E  E 30404 cattatgaaaaaattgttgattttatgaatgcgagaataaatgca 106 H  Y  E  K  I  V  D  F  M  N  A  R  I  N  A 30449 tga 30451 121 * 3AORF041, Nucleotides and amino acids sequences (SEQ ID NO. 22) 21497 atgtttggatttaccaaacgacacgaacaagattggcgtttaacg 1 M  F  G  F  T  K  R  H  E  Q  D  W  R  L  T 21542 cgattagaagaaaatgataagactatgtttgaaaaattcgacaga 16 R  L  E  E  N  D  K  T  M  F  E  K  F  D  R 21587 atagaagacagtctgagaacgcaagaaaaaatttatgacaagtta 31 I  E  D  S  L  R  T  Q  E  K  I  Y  D  K  L 21632 gatagaaatttcgaagaactaaggcgtgacaaagaagaagatgaa 46 D  R  N  F  E  E  L  R  R  D  K  E  E  D  E 21677 aaaaataaagagaaaaatgctaaaaatattagagacatcaagatg 61 K  N  K  E  K  N  A  K  N  I  R  D  I  K  M 21722 tggattctaggattaatagggacgattctaagtacatttgttata 76 W  I  L  G  L  I  G  T  I  L  S  T  F  V  I 21767 gccttgttaaaaactatttttggcatttaa 21796 91 A  L  L  K  T  I  F  G  I  * 3AORF079, Nucleotides and amino acids sequences (SEQ ID NO. 24) 34231 atgcaacatcaagcttatatcaatgcttctgttgacattagaatt 1 M  Q  H  Q  A  Y  I  N  A  S  V  D  I  R  I 34276 cctacagaagtcgaaagtgttaattacaatcagattgataaagaa 16 P  T  E  V  E  S  V  N  Y  N  Q  I  D  K  E 34321 aaagaaaatttggcggactatttatttaataatccaggtgaacta 31 K  E  N  L  A  D  Y  L  F  N  N  P  G  E  L 34366 ttaaaatataacgttataaatattaaggttttagatttagaggtg 46 L  K  Y  N  V  I  N  I  K  V  L  D  L  E  V 34411 gaatga 34416 61 E  * 77ORF001, Nucleotides and amino acids sequences (SEQ ID NO. 26) 8481 atgggagaaagaataaaaggtttatctataggtttggatttagat 1 M  G  E  R  I  K  G  L  S  I  G  L  D  L  D 8526 gcagcaaatttaaatagatcatttgcagaaatcaaacgaaacttt 16 A  A  N  L  N  R  S  F  A  E  I  K  R  N  F 8571 aaaactttaaattctgacttaaaattaacaggcaacaacttcaaa 31 K  T  L  N  S  D  L  K  L  T  G  N  N  F  K 8616 tataccgaaaaatcaactgatagttacaaacaaaggattaaagaa 46 Y  T  E  K  S  T  D  S  Y  K  Q  R  I  K  E 8661 cttgatggaactatcacaggttataagaaaaacgttgatgattta 61 L  D  G  T  I  T  G  Y  K  K  N  V  D  D  L 8706 gccaagcaatatgacaaggtatctcaagaacagggcgaaaacagt 76 A  K  Q  Y  D  K  V  S  Q  E  Q  G  E  N  S 8751 gcagaagctcaaaagttacgacaagaatataacaaacaagcaaat 91 A  E  A  Q  K  L  R  Q  E  Y  N  K  Q  A  N 8796 gagctgaattatttagaaagagaattacaaaaaacatcagccgaa 106 E  L  N  Y  L  E  R  E  L  Q  K  T  S  A  E 8841 tttgaagagttcaaaaaagctcaagttgaagctcaaagaatggca 121 F  E  E  F  K  K  A  Q  V  E  A  Q  R  M  A 8886 gaaagtggctggggaaaaaccagtaaagtttttgaaagtatggga 136 E  S  G  W  G  K  T  S  K  V  F  E  S  M  G 8931 cctaaattaacaaaaatgggtgatggtttaaaatccattggtaaa 151 P  K  L  T  K  M  G  D  G  L  K  S  I  G  K 8976 ggtttgatgattggtgtaactgcacctgttttaggtattgcagca 166 G  L  M  I  G  V  T  A  P  V  L  G  I  A  A 9021 gcatcaggaaaagcttttgcagaagttgataaaggtttagatact 181 A  S  G  K  A  F  A  E  V  D  K  G  L  D  T 9066 gttactcaagcaacaggcgcaacaggcagtgaattaaaaaaattg 196 V  T  Q  A  T  G  A  T  G  S  E  L  K  K  L 9111 cagaactcatttaaagatgtttatggcaattttccagcagatgct 211 Q  N  S  F  K  D  V  Y  G  N  F  P  A  D  A 9156 gaaactgttggtggagttttaggagaagttaatacaaggttaggt 226 E  T  V  G  G  V  L  G  E  V  N  T  R  L  G 9201 tttacaggtaaagaacttgaaaatgccacagagtcattcttgaaa 241 F  T  G  K  E  L  E  N  A  T  E  S  F  L  K 9246 ttcagtcatataacaggttctgacggtgtgcaagccgtacagtta 256 F  S  H  I  T  G  S  D  G  V  Q  A  V  Q  L 9291 attacccgtgcaatgggcgatgcaggtatcgaagcaagtgaatat 271 I  T  R  A  M  G  D  A  G  I  E  A  S  E  Y 9336 caaagtgttttggatatggtagcaaaagcggcgcaagctagtggg 286 Q  S  V  L  D  M  V  A  K  A  A  Q  A  S  G 9381 ataagtgttgatacattagctgatagtattactaaatacggcgct 301 I  S  V  D  T  L  A  D  S  I  T  K  Y  G  A 9426 ccaatgagagctatgggctttgagatgaaagaatcaattgcttta 316 P  M  R  A  M  G  F  E  M  K  E  S  I  A  L 9471 ttctctcaatgggaaaagtcaggcgttaatactgaaatagcattc 331 F  S  Q  W  E  K  S  G  V  N  T  E  I  A  F 9516 agtggtttgaaaaaagctatatcaaattggggtaaagctggtaaa 346 S  G  L  K  K  A  I  S  N  W  G  K  A  G  K 9561 aacccaagagaagaatttaagaagacattagcagaaattgaaaag 361 N  P  R  E  E  F  K  K  T  L  A  E  I  E  K 9606 acgccggatatagctagcgcaacaagtttagcgattgaagcattt 376 T  P  D  I  A  S  A  T  S  L  A  I  E  A  F 9651 ggtgcaaaggcaggtcctgatttagcagacgctattaaaggtggt 391 G  A  K  A  G  P  D  L  A  D  A  I  K  G  G 9696 cgctttagttatcaagaatttttaaaaactattgaagattcccaa 406 R  F  S  Y  Q  E  F  L  K  T  I  E  D  S  Q 9741 ggcacagtaaaccaaacatttaaagattctgaaagtggctccgaa 421 G  T  V  N  Q  T  F  K  D  S  E  S  G  S  E 9786 agatttaaagtagcaatgaataaattaaaattagtaggtgctgat 436 R  F  K  V  A  M  N  K  L  K  L  V  G  A  D 9831 gtatgggcttctattgaaagtgcgtttgctcccgtaatggaagaa 451 V  W  A  S  I  E  S  A  F  A  P  V  M  E  E 9876 ttaatcaaaaagctatctatagcggttgattggttttccaattta 466 L  I  K  K  L  S  I  A  V  D  W  F  S  N  L 9921 agtgatggttctaaaagatcaattgttattttcagtggtattgct 481 S  D  G  S  K  R  S  I  V  I  F  S  G  I  A 9966 gctgcaattggtcctgtagtttttgggttaggtgcatttataagt 496 A  A  I  G  P  V  V  F  G  L  G  A  F  I  S 10011 acaattggcaatgcagtaactgtattagctccattgttagctagt 511 T  I  G  N  A  V  T  V  L  A  P  L  L  A  S 10056 attgcaaaggctggtggattgattagttttttatcgactaaagta 526 I  A  K  A  G  G  L  I  S  F  L  S  T  K  V 10101 cctatattaggaactgtcttcacagctttaactggtccaattggc 541 P  I  L  G  T  V  F  T  A  L  T  G  P  I  G 10146 attgtattaggtgtattggctggtttagcagtcgcatttacaatt 556 I  V  L  G  V  L  A  G  L  A  V  A  F  T  I 10191 gcttataagaaatctgaaacatttagaaattttgttaatggtgca 571 A  Y  K  K  S  E  T  F  R  N  F  V  N  G  A 10236 attgaaagtgttaaacaaacatttagtaattttattcaatttatt 586 I  E  S  V  K  Q  T  F  S  N  F  I  Q  F  I 10281 caacctttcgttgattctgttaaaaacatctttaaacaagcgata 601 Q  P  F  V  D  S  V  K  N  I  F  K  Q  A  I 10326 tcagcaatagttgatttcgcaaaagatatttggagtcaaatcaat 616 S  A  I  V  D  F  A  K  D  I  W  S  Q  I  N 10371 ggattctttaatgaaaacggaatttccattgttcaagcacttcaa 631 G  F  F  N  E  N  G  I  S  I  V  Q  A  L  Q 10416 aatatatgcaactttattaaagcgacatttgaatttattttaaat 646 N  I  C  N  F  I  K  A  I  F  E  F  I  L  N 10461 tttgtaattaaaccaattatgttcgcgatttggcaagtgatgcaa 661 F  V  I  K  R  I  M  F  A  I  W  Q  V  M  Q 10506 tttatttggccggcggttaaagccttgattgtcagtacttgggag 676 F  I  W  P  A  V  K  A  L  I  V  S  T  W  E 10551 aacataaaaggtgtaatacaaggtgctttaaatatcatacttggc 691 N  I  K  G  V  I  Q  G  A  L  N  I  I  L  G 10596 ttgattaagttcttctcaagtttattcgttggtgattggcgagga 706 L  I  K  F  F  S  S  L  F  V  G  D  W  R  G 10641 gtttgggacgccgttgtgatgattcttaaaggagcagttcaatta 721 V  W  D  A  V  V  M  I  L  K  G  A  V  Q  L 10686 atttggaatttagttcaattatggtttgtaggtaaaatacttggt 736 I  W  N  L  V  Q  L  W  F  V  G  K  I  L  G 10731 gttgttaggtactttggcgggttgctaaaaggattgatagcagga 751 V  V  R  Y  F  G  G  L  L  K  G  L  I  A  G 10776 atttgggacgtaataagaagtatattcagtaaatctttatcagca 766 I  W  D  V  I  R  S  I  F  S  K  S  L  S  A 10821 atttggaatgcaacaaaaagtatttttggatttttatttaatagc 781 I  W  N  A  T  K  S  I  F  G  F  L  F  N  S 10866 gtaaaatcaattttcacaaatatgaaaaattggttatctaatact 796 V  K  S  I  F  T  N  M  K  N  W  L  S  N  T 10911 tggagcagtatccgtacgaatacaataggaaaagcgcagtcatta 811 W  S  S  I  R  T  N  T  I  G  K  A  Q  S  L 10956 tttagtggcgtcaaatcaaaatttactaatttatggaatgcgacg 826 F  S  G  V  K  S  K  F  T  N  L  W  N  A  T 11001 aaagaaatttttagtaatttaagaaattggatgtcaaatatttgg 841 K  E  I  F  S  N  L  R  N  W  M  S  N  I  W 11046 aattccattaaagataatacggtaggaattgcaagccgtttatgg 856 N  S  I  K  D  N  T  V  G  I  A  S  R  L  W 11091 agtaaggtacgtggaattttcacaaatatgcgcgatggcttgagt 871 S  K  V  R  G  I  F  T  N  M  R  D  G  L  S 11136 tccattatagataagattaaaagtcatatcggcggtatggtaagc 886 S  I  I  D  K  I  K  S  H  I  G  G  M  V  S 11181 gctattaaaaaaggacttaataaattaatcgacggtttaaactgg 901 A  I  K  K  G  L  N  K  L  I  D  G  L  N  W 11226 gtcggtggtaagttgggaatggataaaatacctaagttacacact 916 V  G  G  K  L  G  M  D  K  I  P  K  L  H  T 11271 ggtacagagcacacacatactactacaagattagttaagaacggt 931 G  T  E  H  T  H  T  T  T  R  L  V  K  N  G 11316 aagattgcacgtgacacattcgctacagttggggataagggacgc 946 K  I  A  R  D  T  F  A  T  V  G  D  K  G  R 11361 ggaaatggtccaaatggttttagaaatgaaatgattgaattccct 961 G  N  G  P  N  G  F  R  N  E  M  I  E  F  P 11406 aacggtaaacgtgtaatcacacctaatacagatactaccgcttat 976 N  G  K  R  V  I  T  P  N  T  D  T  T  A  Y 11451 ttacctaaaggctcaaaagtatacaacggtgcacaaacttattca 991 L  P  K  G  S  K  V  Y  N  G  A  Q  T  Y  S 11496 atgttaaacggaacgcttccaagatttagtttaggtactatgtgg 1006 M  L  N  G  T  L  P  R  F  S  L  G  T  M  W 11541 aaagatattaaatctggtgcatcatcggcatttaactggacaaaa 1021 K  D  I  K  S  G  A  S  S  A  F  N  W  T  K 11586 gataaaataggtaaaggtaccaaatggcttggcgataaagttggc 1036 D  K  I  G  K  G  T  K  W  L  G  D  K  V  G 11631 gatgttttagattttatggaaaatccaggcaaacttttaaattat 1051 D  V  L  D  F  M  E  N  P  G  K  L  L  N  Y 11676 atacttgaagcttttggaattgatttcaattctttaactaaaggt 1066 I  L  E  A  F  G  I  D  F  N  S  L  T  K  G 11721 atgggaattgcaggcgacataacaaaagctgcatggtctaagatt 1081 M  G  I  A  G  D  I  T  K  A  A  W  S  K  I 11766 aagaaaagtgctactgattggataaaagaaaatttagaagctatg 1096 K  K  S  A  T  D  W  I  K  E  N  L  E  A  M 11811 ggcggtggcgatttagtcggcggaatattagaccctgacaaaatt 1111 G  G  G  D  L  V  G  G  I  L  D  P  D  K  I 11856 aattatcattatggacgtaccgcagcttataccgctgcaactgga 1126 N  Y  H  Y  G  R  T  A  A  Y  T  A  A  T  G 11901 agaccatttcatgaaggtgtcgattttccatttgtatatcaagaa 1141 R  P  F  H  E  G  V  D  F  P  F  V  Y  Q  E 11946 gttagaacgccgatgggtggcagacttacaagaatgccatttatg 1156 V  R  T  P  M  G  G  R  L  T  R  M  P  F  M 11991 tctggtggttatggtaattatgtaaaaattactagtggcgttatc 1171 S  G  G  Y  G  N  Y  V  K  I  T  S  G  V  I 12036 gatatgctatttgcgcatttgaaaaactttagcaaatcaccacct 1186 D  M  L  F  A  H  L  K  N  F  S  K  S  P  P 12081 agtggcacgatggtaaagcccggtgatgttgttggtttaactggt 1201 S  G  T  M  V  K  P  G  D  V  V  G  L  T  G 12126 aataccggatttagtacaggaccacatttacattttgaaatgagg 1216 N  T  G  F  S  T  G  P  H  L  H  F  E  M  R 12171 agaaatggacgacattttgaccctgaaccatatttaaggaatgct 1231 R  N  G  R  H  F  D  P  E  P  Y  L  R  N  A 12216 aagaaaaaaggaagattatcaataggtggtggcggtgctacttct 1246 K  K  K  G  R  L  S  I  G  G  G  G  A  T  S 12261 ggaagtggcgcaacttatgccagtcgagtaatccgacaagcgcaa 1261 G  S  G  A  T  Y  A  S  R  V  I  R  Q  A  Q 12306 agtattttaggtggtcgttataaaggtaaatggattcatgaccaa 1276 S  I  L  G  G  R  Y  K  G  K  W  I  H  D  Q 12351 atgatgcgcgttgcaaaacgtgaaagtaactaccagtcaaatgca 1291 M  M  R  V  A  K  R  E  S  N  Y  Q  S  N  A 12396 gtgaataactgggatataaatgctcaaagaggagacccatcaaga 1306 V  N  N  W  D  I  N  A  Q  R  G  D  P  S  R 12441 ggattattccaaatcatcggctcaacttttagagcaaacgctaaa 1321 G  L  F  Q  I  I  G  S  T  F  R  A  N  A  K 12486 cgtggatatactaactttaataatccagtacatcaaggtatctca 1336 R  G  Y  T  N  F  N  N  P  V  H  Q  G  I  S 12531 gcaatgcagtacattgttagacgatatggttggggtggttttaaa 1351 A  M  Q  Y  I  V  R  R  Y  G  W  G  G  F  K 12576 cgtgctggtgattacgcatatgctacaggtggaaaagtttttgat 1366 R  A  G  D  Y  A  Y  A  T  G  G  K  V  F  D 12621 ggttggtataacttaggtgaagacggtcatccagaatggattatt 1381 G  W  Y  N  L  G  E  D  G  H  P  E  W  I  I 12666 ccaacagatccagctcgtagaaatgatgcaatgaagattttgcat 1396 P  T  D  P  A  R  R  N  D  A  M  K  I  L  H 12711 tatgcagcagcagaagtaagagggaaaaaagcgagtaaaaataag 1411 Y  A  A  A  E  V  R  G  K  K  A  S  K  N  K 12756 cgtcctagccaattatcagacttaaacgggtttgatgatcctagc 1426 R  P  S  Q  L  S  D  L  N  G  F  D  D  P  S 12801 ttattattgaaaatgattgaacaacagcaacaacaaatagcttta 1441 L  L  L  K  M  I  E  Q  Q  Q  Q  Q  I  A  L 12846 ttactgaaaatagcacaatctaacgatgtgattgcagataaagat 1456 L  L  K  I  A  Q  S  N  D  V  I  A  D  K  D 12891 tatcagccgattattgacgaatacgcttttgataaaaaggtgaac 1471 Y  Q  P  I  I  D  E  Y  A  F  D  K  K  V  N 12936 gcgtctatagaaaagcgagaaaggcaagaatcaacaaaagtaaag 1486 A  S  I  E  K  R  E  R  Q  E  S  T  K  V  K 12981 tttagaaaaggaggaattgctattcaatga 13010 1501 F  R  K  G  G  I  A  I  Q  * 96ORF048, Nucleotides and amino acids sequences (SEQ ID NO. 28) 4952 atgtattacaaaattggtgagataaaaaacaaaattataagcttt 1 M  Y  Y  K  I  G  E  I  K  N  K  I  I  S  F 4997 aacgggtttgaatttaaagtgtctgtgatgaagagacatgacggt 16 N  G  F  E  F  K  V  S  V  M  K  R  H  D  G 5042 atcagtatacaaatcaaggatatgaataatgttccacttaaatcg 31 I  S  I  Q  I  K  D  M  N  N  V  P  L  K  S 5087 tttcatgtcatagatttaagcgaactatatattgcgacggatgca 46 F  H  V  I  D  L  S  E  L  Y  I  A  T  D  A 5132 atgcgtgacgttataaacgaatggattgaaaataacacagatgaa 61 M  R  D  V  I  N  E  W  I  E  N  N  T  D  E 5177 caggacaaactaattaacttagtcatgaaatggtag 5212 76 Q  D  K  L  I  N  L  V  M  K  W  * 96ORF078, Nucleotides and amino acids sequences (SEQ ID NO. 30) 10148 atgaatataatgcaattcaaaagcttattgaaatcgatgtatgaa 1 M  N  I  M  Q  F  K  S  L  L  K  S  M  Y  E 10193 gagacaaagcaaagcgacccgattgtagcaaatgtatatatcgag 16 E  T  K  Q  S  D  P  I  V  A  N  V  Y  I  E 10238 actggttgggcggtcaatagattgttggacaataacgagttatcg 31 T  G  W  A  V  N  R  L  L  D  N  N  E  L  S 10283 cctttcgatgattacgacagagttgaaaagaaaatcatgaatgaa 46 P  F  D  D  Y  D  R  V  E  K  K  I  M  N  E 10328 atcaactggaagaaaacacacattaaggagtgttaa 10363 61 I  N  W  K  K  T  H  I  K  E  C  * 96ORF100, Nucleotides and amino acids sequences (SEQ ID NO. 32) 11008 atgcaacaacaagcatatataaacgcaacaattgatataagaata 1 M  Q  Q  Q  A  Y  I  N  A  T  I  D  I  R  I 11053 cctacagaagttgaatatcagcattacgatgatgtggataaagaa 16 P  T  E  V  E  Y  Q  H  Y  D  D  V  D  K  E 11098 aaagatacgctggcaaagcgcttagatgacaatccggacgaatta 31 K  D  T  L  A  K  R  L  D  D  N  P  D  E  L 11143 ctaaagtatgacaacataacaataagacatgcatatatagaggtg 46 L  K  Y  D  N  I  T  I  R  H  A  Y  I  E  V 11188 gaataa 11193 61 E  *

TABLE 4 Similarities with public sequences Query = pt|100214 3AORF033 3A_NT|30089-30451|2 1 (120 letters) Database: nr 445,337 sequences; 137,034,979 total letters Score E Sequences producing significant alignments: (bits) Value gi|246049|bbs|83873 neurofilament protein M [rats, Peptide Part . . . 35 0.16 gi|56752|emb|CAA78136|(Z12152) Neurofilament protein middle (N . . . 35 0.16 gi|128150|sp|P12839|NFM_RAT NEUROFILAMENT TRIPLET M PROTEIN (16 . . . 35 0.16 gi|482393|pir||A45669 neurofilament triplet M protein - rat > gi . . . 35 0.16 gi|6587836|gb|AAF18525.1|AC006551_11 (AC006551) Unknown protein . . . 35 0.16 gi|2459888 (AF005844) anon1A3 [Drosophila yakuba] 35 0.21 gi|1621107 (U62026) cardiac muscle factor 1 CMF1 [Gallus gallus] 34 0.27 gi|160409 (M69183) mature-parasite-infected erythrocyte surface . . . 34 0.36 gi|3044185 (AF056936) mature parasite-infected erythrocyte surf . . . 34 0.36 gi|323126|pir||A45605 mature-parasite-infected erythrocyte surf . . . 34 0.36 gi|482391|pir||A45555 glutamate rich protein - Plasmodium falci . . . 34 0.47 gi|3413892|dbj|BAA32310| (AB007934) KIAA0465 protein [Homo sapi . . . 33 0.61 gi|6273778|gb|AAF06360.1|AF141968_1 (AF141968) trabeculin-alpha . . . 33 0.61 gi|5821434|dbj|BAA83821.1| (AB029290) actin binding protein ABP . . . 33 0.61 Query = pt|100214 3AORF033 3A_NT|30089-30451|2 1 (120 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|P12839 NFN_RAT NEUROFILAMENT TRIPLET M PROTEIN (160 KD NEUR . . . 35  0.040 sp|Q02555 RNT1_YEAST RIBONUCLEASE III (EC 3.1.26.3) (RNASE III) . 32 0.34 sp|P32841 R114_YEAST MEIOTIC RECOMBINATION PROTEIN REC114. 32 0.34 sp|P29681 IMP2_DROME 20-HYDROXYECDYSONE PROTEIN PRECURSOR (20- . . . 32 0.45 sp|O00294 TUL1_HUMAN TUBBY RELATED PROTEIN 1 (TUBBY-LIKE PROTE . . . 32 0.45 sp|P28608 DNAK_BORBU DNAK PROTEIN (HEAT SHOCK PROTEIN 70) (HSP . . . 31 0.77 sp|Q57639 Y175_METJA HYPOTHETICAL PROTEIN MJ0175. 31 0.77 Query = pt|100222 3AORF041 3A_NT|21497-21796|2 1 (99 letters) Database: nr 445,337 sequences; 137,034,979 total letters Score E Sequences producing significant alignments: (bits) Value gi|6382413|gb|AAF07723.1|AE001584_20 (AE001584) conserved hypot . . . 30 6.6 gi|130509|sp|P29152|POLG_PSBMV GENOME POLYPROTEIN (CONTAINS: N- . . . 29 8.7 gi|5104896|dbj|BAA80210.1| (AP000061) 356aa long hypothetical t . . . 29 8.7 Query = pt|100222 3AORF041 3A_NT|21497-21796|2 1 (99 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|P29152 POLG_PSBMV GENOME POLYPROTEIN [CONTAINS: N-TERMINAL . . . 29 2.1 sp|P54470 YQFL_BACSU HYPOTHETICAL 30.3 KD PROTEIN IN GLYS-DNAG . . . 29 2.8 Query = pt|100260 3AORF079 3A_NT|34231-34416|1 1 (61 letters) Database: nr 445,337 sequences; 137,034,979 total letters Score E Sequences producing significant alignments: (bits) Value gi|2496354|sp|P75400|Y264_MYCPN HYPOTHETICAL PROTEIN MG264 HOMO . . . 29 3.7 gi|6175671|gb|AAF05141.1|AF162221_27 (AF162221) ORF27 [Xestia c . . . 29 4.8 gi|6136641|sp|O78467|YCF4_GUITH HYPOTHETICAL 20.9 KD PROTEIN YC . . . 29 6.3 gi|2621735 (AE000845) conserved protein [Methanobacterium therm . . . 28 8.3 gi|3845294 (AE001421) rRNA methylase (SpoU family) (OO, TP) [Pl . . . 28 8.3 Query = pt|100260 3AORF079 3A_NT|34231-34416|1 1 (61 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|P75400 Y264_MYCPN HYPOTHETICAL PROTEIN MG264 HOMOLOG. 29  0.83 sp|078467 YCF4_GUITH HYPOTHETICAL 20.9 KD PROTEIN YCF4. 29 1.4 sp|P30619 SEC1_YEAST PROTEIN TRANSPORT PROTEIN SEC1. 27 3.2 sp|P43055 YLI1_MYCHO HYPOTHETICAL 59.8 KD PROTEIN IN LICA 3′RE . . . 27 4.2 sp|O62757 CSF2_FELCA GRANULOCYTE-MACROPHAGE COLONY-STIMULATING . . . 27 5.5 sp|P35725 YKG3_YEAST HYPOTHETICAL 19.0 KD PROTEIN IN MNR2-MSN4 . . . 26 7.2 sp|P10942 YHA2_CRYPA HYPOTHETICAL PROTEIN 2 IN HYPOVIRULENCE-A . . . 26 9.5 sp|P48749 CSF2_CANFA GRANULOCYTE-MACROPHAGE COLONY-STIMULATING . . . 26 9.5 sp|P54679 PMA1_DICDI PROBABLE PLASMA MEMBRANE ATPASE (EC 3.6.1 . . . 26 9.5 Query = pt|100001 77ORF001 77_NT|8481-13010|3 1 (1509 letters) Database: nr 445,337 sequences; 137,034,979 total letters Searching.................................................done Score E Sequences producing significant alignments: (bits) Value gi|3341923|dbj|BAA31889.1)|(AB009866) orf 16 [bacteriophage phi . . . 797 0.0 gi|3341922|dbj|BAA31888.1)|(AB009866) orf 15 [bacteriophage phi . . . 268 3e-70 gi|3341924|dbj|BAA31890.1)|(AB009866) orf 17 [bacteriophage phi . . . 234 4e-60 gi|2392838 (AF011378) unknown [Bacteriophage sk1] 150 8e-35 gi|3282276 (AF009630) 116 [bacteriophage bIL170] 131 6e-29 gi|4530151|gb|AAD21891.1|(AF085222) putative tail component pr . . . 126 1e-27 gi|2935689|gb|AAC39295.1|(AF115102) orf1626 gp [Streptococcus . . . 116 1e-24 gi|1926360|emb|CAA66745|(X98106) minor capsid protein [Bacteri . . . 106 2e-21 gi|2935674|gb|AAC39281.1|(AF115103) orf1560 gp [Streptococcus . . .  98 5e-19 gi|4530152|gb|AAD21892.1|(AF085222) putative tail component pr . . .  96 2e-18 gi|1722872|sp|P54334|XKDO_BACSU PHAGE-LIKE ELEMENT PBSX PROTEIN . . .  83 2e-14 gi|2764873|emb|CAA66557|(X97918) gene 18.1 [Bacteriophage SPP1]  78 7e-13 gi|1353559 (U38906) ORF42 [Bacteriophage rlt]  78 7e-13 gi|1176754|sp|P45931|YQBO_BACSU HYPOTHETICAL 171.0 KD PROTEIN I . . .  77 9e-13 gi|2313617|gb|AAD07571.1|(AE000565) conserved hypothetical sec . . .  77 1e-12 gi|4154996 (AE001480) putative Outer membrane protein [Helicoba . . .  75 3e-12 gi|2688140 (AE001134) B. burgdorferi predicted coding region BB . . .  71 5e-11 gi|6599346|emb|CAB63691.1| (AJ251790) hypothetical protein [Lac . . .  70 1e-10 gi|1073751|pir||JC2569 tagE protein - Vibrio cholerae (strain 0 . . .  70 2e-10 gi|2688203 (AE001137) conserved hypothetical protein [Borrelia . . .  70 2e-10 gi|3860964|emb|CAA14864|](AJ235271) unknown [Rickettsia prowaze . . .  70 2e-10 gi|623073 (L02496) unknown protein [Bacteriophage LL-H]  69 3e-10 gi|4980914|gb|AAD35494.1|AE001720_8 (AE001720) conserved hypoth . . .  69 3e-10 gi|1175836|sp|P44693|YEBA_HAEIN HYPOTHETICAL PPOTEIN HI0409 > gi . . .  68 8e-10 gi|1944592|emb|CAB08078|(Z94121) hypothetical protein Rv3896c . . .  65 4e-09 gi|6136204|sp|O64220|VG26_BPMD2 MINOR TAIL PROTEIN GP26 > gi|317 . . .  65 4e-09 gi|1369948|emb|CAA59194| (X84706) host interacting protein [Bac . . .  63 1e-08 gi|3947462|emb|CAA07113.1| (AJ006589) gp43 [Bacteriophage phi-C31]  63 1e-08 gi|2444119 (U88974) ORF40 [Streptococcus thermophilus temperate . . .  62 3e-08 gi|4336054|gb|AAD17585|(AF068845) gp17 [Mycobacteriophage TM4]  61 1e-07 gi|6460534|gb|AAF12240.1|AE001862_66 (AE001862) minor tail prot . . .  61 1e-07 gi|3287732|sp|O05156|ALE1_STACP GLYCYL-GLYCINE ENDOPEPTIDASE AL . . .  59 4e-07 gi|6137045|emb|CAB59600.1|(AL132662) possible peptidase [Strep . . .  58 5e-07 gi|79926|pir||A25881 lysostaphin precursor - Staphylococcus sim . . .  58 7e-07 gi|126496|sp|P10548|LSTP_STAST LYSOSTAPHIN PRECURSOR (GLYCYL-GL . . .  58 7e-07 gi|3287967|sp|P10547|LSTP_STASI LYSOSTAPHIN PRECURSOR (GLYCYL-G . . .  58 7e-07 qi|5042257|emb|CAB44511.1| (AL078618) hypothetical protein [Str . . .  57 1e-06 Query = pt|100001 77ORF001 77_NT|8481-13010|3 1 (1509 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|P54334 XKDO_BACSU PHAGE-LIKE ELEMENT PBSX PROTEIN XKDO. 83 5e-15 sp|P45931 YQBO_BACSU HYPOTHETICAL 171.0 KD PROTEIN IN SPOIIIC- . . . 77 2e-13 sp|P44693 YEBA_HAEIN HYPOTHETICAL PROTEIN HI0409. 68 2e-10 sp|O64220 VG26_BPMD2 MINOR TAIL PROTEIN GP26. 65 9e-10 sp|O05156 ALE1_STACP GLYCYL-GLYCINE ENDOPEPTIDASE ALE-1 PRECUR . . . 59 9e-08 sp|P10547 LSTP_STASI LYSOSTAPHIN PRECURSOR (EC 3.4.24.75) (GLY . . . 58 2e-07 sp|P10548 LSTP_STAST LYSOSTAPHIN PRECURSOR (EC 3.4.24.75) (GLY . . . 58 2e-07 sp|P24204 YEBA_ECOLI HYPOTHETICAL 46.7 KD PROTEIN IN MSBB-RUVB . . . 55 1e-06 sp|P51731 YO27_BPHP1 HYPOTHETICAL 72.8 KD PROTEIN IN LYS 3′REG . . . 55 1e-06 sp|Q09857 YAF3_SCHPO HYPOTHETICAL 118.6 KD PROTEIN C29E6.03C I . . . 51 2e-05 sp|QC5233 VG26_BPML5 MINOR TAIL PROTEIN GP26. 47 2e-04 sp|P39922 MYS3_HYDAT MYOSIN HEAVY CHAIN, CLONE 203 (FRAGMENT) . . . 47 3e-04 sp|P12844 MYSA_CAEEL MYCSIN HEAVY CHAIN A (MHC A) . . . 47 3e-04 sp|P12845 MYSC_CAEEL MYCSIN HEAVY CHAIN C (MHC C) . . . 46 5e-04 sp|P37690 YIBP_ECOLI HYPOTHETICAL 46.6 KD PROTEIN IN SECB-TDH . . . 46 5e-04 sp|P24733 MYS_AEQIR MYOSIN HEAVY CHAIN, STRIATED MUSCLE. 45 9e-04 Query = pt|100405 96ORF048 96_NT|4952-5212|1 1 (86 letters) Database: nr 445,337 sequences; 137,034,979 total letters Score E Sequences producing significant alignments: (bits) Value gi|3341947|dbj|BAA31913.1| (AB009866) orf 39 [bacteriophage phi . . . 116  4e-26 gi|3183240|sp|Q58352|Y942_METJA PROBABLE ATP-DEPENDENT HELICASE . . . 31 2.7 gi|4033401|sp|P94281|GYRB_BARBA DNA GYRASE SUBUNIT B > gi|176606 . . . 30 4.6 gi|3258109|dbj|BAA30792|(AP000006) 320aa long hypothetical pro . . . 29 6.1 gi|5457925|emb|CAB49415.1| (AJ248284) hypothetical protein [Pyr . . . 29 6.1 gi|4678268|emb|CAB41176.1| (AL049660) putative protein [Arabido . . . 29 8.0 Query = pt|100405 96ORF048 96_NT|4952-5212|1 1 (86 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|Q58352 Y942_METJA PROBABLE ATP-DEPENDENT HELICASE MJ0942. 31  0.69 sp|P94281 GYRB_BARBA DNA GYRASE SUBUNIT B (EC 5.99.1.3). 30 1.2 sp|Q60384 Y077_METJA HYPOTHETICAL PROTEIN MJ0077. 29 2.7 sp|Q03164 HRX_HUMAN ZINC FINGER PROTEIN HRX (ALL-1) (TRITHORAX . . . 28 3.5 sp|P55200 HRX_MOUSE ZINC FINGER PROTEIN HRX (ALL-1) (FRAGMENT). 28 3.5 sp|Q01926 MRS2_YEAST MITOCHONDRIAL RNA SPLICING PROTEIN MRS2 P . . . 27 6.0 sp|P47508 SYL_MYCGE LEUCYL-TRNA SYNTHETASE (EC 6.1.1.4) (LEUCI . . . 27 6.0 sp|P14933 YP60_METTM HYPOTHETICAL 60.5 KD PROTEIN. 27 7.9 Query = pt|100435 96ORF078 96_NT|10148-10363|1 1 (71 letters) Database: nr 445,337 sequences; 137,034,979 total letters Score E Sequences producing significant alignments: (bits) Value gi|167324 (M92051) 5′ start site is putative; putative [Gossypi . . . 30 2.1 gi|3875068|emb|CAB03979.1| (Z81485) cDNA EST EMBL:T02038 comes . . . 30 2.1 gi|232024|sp|Q01197|E6_GOSHI PROTEIN E6 > gi|421806|pir||A46130 . . . 30 2.1 gi|2129495|pir||S65063 fiber protein E6 (clone SIE6-2A) - sea-i . . . 30 2.1 gi|2982648|emb|CAA05305| (AJ002294) penicillin-binding protein . . . 30 2.7 gi|4033461|sp|O51889|REP_BUCAP ATP-DEPENDENT DNA HELICASE REP > . . . 29 3.5 Query = pt|100435 96ORF078 96_NT|10148-10363|1 1 (71 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|Q01197 E6_GOSHI PROTEIN E6. 30  0.57 sp|O51889 REP_BUCAP ATP-DEPENDENT DNA HELICASE REP (EC 3.6.1.-). 29  0.97 sp|P53125 YGN3_YEAST HYPOTHETICAL 145.6 KD PROTEIN IN RPL1B-CE . . . 28 2.9 sp|P03459 HEMA_IAFPR HEMAGGLUTININ PRECURSOR [CONTAINS: HEMAGG . . . 28 2.9 Query = pt|100457 96ORF100 96_NT|11008-11193|3 1 (61 letters) Database: nr 445,337 sequences; 137,034,979 total letters Score E Sequences producing significant alignments: (bits) Value gi|6687541|emb|CAB65007.1| (Y17316) transmembrane protein [Erys . . . 29 3.7 gi|6175777|gb|AAF05247.1|AF162221_133 (AF162221) ORF133 [Xestia . . . 29 6.3 gi|4508013|ref|NP_003445.1||zinc finger protein 200 > gi|622650 . . . 28 8.3 Query = pt|100457 96ORF100 96_NT|11008-11193|3 1 (61 letters) Database: swissprot 83,367 sequences; 30,300,539 total letters Score E Sequences producing significant alignments: (bits) Value sp|P98182 Z200_HUMAN ZINC FINGER PROTEIN ZNF200. 28 1.9 sp|Q08014 MEDB_GIALA MEDIAN BODY PROTEIN. 28 2.5 sp|P18247 POLG_PVYN GENOME POLYPROTEIN [CONTAINS: N-TERMINAL P . . . 27 4.2 sp|P75211 P200_MYCPN PROTEIN P200. 27 4.2 sp|Q02963 POLG_PVYHU GENOME POLYPROTEIN [CONTAINS: N-TERMINAL . . . 27 4.2 sp|P43864 LON_HAEIN ATP-DEPENDENT PROTEASE LA (EC 3.4.21.53). 26 7.2 sp|P54784 ORC1_YEAST ORIGIN RECOGNITION COMPLEX SUBUNIT 1 (ORI . . . 26 7.2

TABLE 5 Optimal global alignment Sequence 1: 3AORF079 Sequence 2: 96ORF100 Substitution matrix: blosum62 Gap penalty: - (11 + 1 * (gap length)) Identical: 37/61 (0.61) Similar:   47/61 (0.77) Score:     181

Sequence 1: 96ORF048 Sequence 2: 77ORF043 Substitution matrix: blosum62 Gap penalty: - (11 + 1 * (gap length)) Identical: 53/86 (0.62) Similar:   68/86 (0.79) Score:     287

Sequence 1: 96ORF048 Sequence 2: 77ORF182 Substitution matrix: blosum62 Gap penalty: - (11 + 1 * (gap length)) Identical: 53/98 (0.54) Similar:   68/98 (0.69) Score:     264

36 1 21 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 1 aattctcgag taaaataaca t 21 2 37 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 2 cgggatccgc ctccttttct caacagtcac ctgattt 37 3 27 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 3 cgggatccat gaggggttcc gaagacg 27 4 24 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 4 cccaagctta caatttggac tttc 24 5 23 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 5 tgagaaaagg aggcggatcc atg 23 6 14 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 6 agctgtcgac gcgt 14 7 14 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 7 agctacgcgt cgac 14 8 17 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 8 taagctgtcg acgcgta 17 9 53 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 9 gatcccggtc gaccaagctt tacccatacg acgtcccaga ctacgccagc tga 53 10 53 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 10 agcttcagct ggcgtagtct gggacgtcgt atgggtaaag cttggtcgac cgg 53 11 27 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 11 ccgctcgagc tccaaattcc aaaacag 27 12 26 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 12 cgggatccaa taagactcct ttttac 26 13 21 DNA Artificial Sequence Description of Artificial Sequence Synthetic oligonucleotide 13 aggagtctta ttggatccat g 21 14 20 DNA Artificial Sequence Description of Artificial Sequence Primer 14 tattatccaa aacttgaaca 20 15 20 DNA Artificial Sequence Description of Artificial Sequence Primer 15 cggtggtata tccagtgatt 20 16 100 PRT Staphylococcus aureus 16 Met Val Asn Val Asp Asn Ala Pro Glu Glu Lys Gly Gln Ala Tyr Thr 1 5 10 15 Glu Met Leu Gln Leu Phe Asn Lys Leu Ile Gln Trp Asn Pro Ala Tyr 20 25 30 Thr Phe Asp Asn Ala Ile Asn Leu Leu Ser Ala Cys Gln Gln Leu Leu 35 40 45 Leu Asn Tyr Asn Ser Ser Val Val Gln Phe Leu Asn Asp Glu Leu Asn 50 55 60 Asn Glu Thr Lys Pro Glu Ser Ile Leu Ser Tyr Ile Ala Gly Asp Asp 65 70 75 80 Pro Ile Glu Gln Trp Asn Met His Lys Gly Phe Tyr Glu Thr Tyr Asn 85 90 95 Val Tyr Val Phe 100 17 43095 DNA Staphylococcus bacteriophage 17 tttaaataaa attttatgcc cccctgccca tcggcttaaa atgttttttc gccgggtacc 60 ggagaggccc aaacgctagc aacgcggata aatttttcat gaaagggggt ctttatatga 120 agttaacaaa aaaacagcta aaagaatata tagaagatta caaaaaatct gatgacatat 180 taattaattt gtatatagaa acatatgaat tttattgtcg gttaagagat gaacttaaaa 240 atagtgattt aatgatagag catacaaaca aggctggtgc gagcaatatt attaagaatc 300 cattaagcat agaactgaca aaaacagttc aaacactaaa taacttactc aagtctatgg 360 gtttaactgc agcacaaaga aaaaagatag ttcaagaaga aggtggattc ggtgactatt 420 aaagttttaa atgaaccttc accaaaacta ttaacaacat ggtatgcaga gcaagtcact 480 caagggaaaa taaaaacaag caaatatgtt agaaaagaat gtgagagaca tcttagatat 540 ctagaaaatg gaggtaaatg ggtatttgat gaagaattag cgcatcgtcc tattcgattt 600 atagaaaagt tttgtaaacc ttccaaagga tctaaacgtc aacttgtatt acagccatgg 660 caacatttta ttatcggcag tttgtttggt tgggttcata aagaaacaaa actgcgcagg 720 tttaaagaag ctttgatatt tatggggcga aaaaatggta aaacaaccac tatttctggg 780 gttgctaact atgctgtatc acaagatgga gaaaatggtg cagaaattca tttgttagca 840 aacgtaatga aacaagctag gattctattt gatgaatcta aggcgatgat taaagctagc 900 ccaaagcttg ataaaaattt cagaacatta agagatgaaa tccattatga cgcaacgata 960 tcaaaaatta tgccccaagc atcagatagc gataagttag atggattgaa tacacacatg 1020 gggatttttg atgaaattca tgaatttaaa gactataaat tgatttcagt tataaaaaac 1080 tcaagagctg caaggttaca acctcttctc atctacatta cgacagcagg gtatcaatta 1140 gatggtccac ttgttgatat ggtagaagcg ggaagagaca ccttagatca aatcatagaa 1200 gacgaaagaa ctttttatta tttagcatct ttggatgatg acgatgatat taatgattcg 1260 tcgaactgga taaaagcaaa tcccaactta ggtgtctcta taaatttaga tgagatgaaa 1320 gaagagtggg aaaaagctaa gagaacacca gctgaacgtg gagattttat aaccaaaagg 1380 tttaatatct ttgctaataa tgacgagatg agttttattg attacccaac actccaaaaa 1440 aataatgaaa ttgtttcttt agaagagctg gaaggcagac cgtgcacgat tggttatgat 1500 ttatcagaaa cagaggactt tacagccgcg tgtgctactt ttgcgttaga taatggtaaa 1560 gttgcagttt tatcgcattc atggattcct aagcacaaag ttgaatattc taacgaaaaa 1620 ataccctata gagaatggga agaagatggc ttattaacag tgcaagataa gccttatatt 1680 gactaccaag atgttttaaa ttggataatt aagatgaatg agcattatgt agtagaaaaa 1740 attacttatg atagagcgaa cgcattcaaa ctaaatcaag agttaaaaaa ttacgggttt 1800 gaaacggaag aaacaagaca aggagctttg accttgagcc ctgcattgaa ggatttaaaa 1860 gaaatgtttt tagatgggaa aataatattt aataataatc ctttaatgaa atggtatatc 1920 aataatgttc agttgaaact agacagaaac ggaaactggt tgccgtctaa gcaaagcaga 1980 tatcgtaaaa tagatggctt tgcagcattt ttaaacacat atacagatat tatgaataaa 2040 gttgtttctg atagtggtga aggaaacata gagtttatta gtattaaaga cataatgcgt 2100 taaggaggtg aatgttatcg caaaagagaa tattgtcaca cgcataaaga aaaaattgat 2160 agacaattgg attgatcagt caacttctaa gctttatgac tttagcccat ggaaaaatag 2220 atctttttgg ggtgtaatta ataatacgct tgaaactaat gaaacgatat tttcagctat 2280 tacaaagtta tctaattcga tggctagttt gcccttgaaa atgtatgaag attataaagt 2340 agttaataca gaagtatctg atttacttac agtgtcaccg aataattctc tgagcagttt 2400 tgattttatt aatcaaattg aaacaatcag aaatgaaaaa ggtaatgcat atgtgctaat 2460 tgaacgagac atctatcatc aaccatcaaa gcttttctta ttaaatccag atgttgttga 2520 aatgttaatt gaaaaccaat cacgtgaact ttattattcc attcatgctg caactggaaa 2580 taaattgatt gttcataata tggacatgtt gcattttaaa cacatcgtgg catctaatat 2640 ggtgcaaggc attagtccga ttgatgtgtt gaagaataca actgattttg ataatgcagt 2700 aagaaccttt aatcttacag aaatgcaaaa acctgattct ttcatgctta aatatggttc 2760 caatgtaggt aaagaaaaaa ggcagcaagt gttagaagat ttcaaacagt actatgaaga 2820 aaacggtgga atattattcc aagagcctgg tgttgaaatc gaaccgttac ctaaaaaata 2880 tgtctctgaa gatatagtgg caagcgagaa tttaacaaga gaaagagtag ctaacgtttt 2940 tcaattgccc tcagtattct taaatgcaag atcaaataca aatttcgcga aaaatgaaga 3000 gttaaacaga ttttacttgc agcatacctt attgccaatc gtcaaacagt atgaagaaga 3060 atttaatcgg aaactactta ctaaaacaga cagagaaaaa aataggtatt ttaaatttaa 3120 cgttaaatct tatttaaggg ctgatagtgc aacacaagca gaagtgtact ttaaagcagt 3180 tcgtagtggt tactacacta taaatgacat tagagagtgg gaagatttac caccagttga 3240 aggtggagat aagccgctaa taagcggtga tttataccca attgacacgc cacttgaatt 3300 aagaaaatct ttgaaaggtg gtgataaaaa tgtcaatgaa agctaagtat tttcaaatga 3360 aaagaaaatc aaaaagtaaa ggtgaaatat ttatttatgg tgatattgta agtgataaat 3420 ggtttgaaag tgatgtaact gctacagatt tcaaaaataa actagatgaa ctaggagaca 3480 tcagtgaaat agatgttcat ataaattcat ctggaggcag tgtatttgaa gggcatgcaa 3540 tatacaatat gctaaaaatg catcctgcaa aaattaatat ctatgtcgat gccttagcgg 3600 catcaattgc tagtgttatc gctatgagtg gtgacactat ttttatgcac aaaaatagtt 3660 ttttaatgat tcataattca tgggttatga ctgtaggtaa tgcagaagag ttaagaaaga 3720 cagcggattt acttgaaaaa acagatgctg ttagtaattc agcttattta gataaagcaa 3780 aagatttaga tcaagaacac ttaaaacaga tgttagatgc agaaacttgg cttactgcag 3840 aagaagcctt gtctttcggc ttgatagatg aaattttagg agctaatgaa ataactgcta 3900 gtatctctaa agagcaatat aagcgtttcg agaacgtccc agaagattta aagaaagatg 3960 tagacaaaat cactaaaatc gatgatgtag atacgtttga attggttgaa acacctaaag 4020 aaagtatgtc actagaagaa aaagaaaaaa gagaaaaaat taaacgcgaa tgcgaaattt 4080 taaaaatgac aatgagttat taggaggaaa tgaaatgccg acattatatg aattaaaaca 4140 atccttaggt atgattggac aacaattaaa aaataaaaat gatgaattga gtcagaaagc 4200 aacagaccca aatattgata tggaagacat caaacaacta gaaacagaaa aagcaggctt 4260 acaacaaaga tttaacattg ttgaaagaca agtaaaagac attgaagaaa aagaaaaagc 4320 gaaagttaaa gacacaggag aagcttatca atctttaaat gatcatgaga agatggttaa 4380 agctaaggca gagttttatc gtcacgcgat tttaccaaat gaatttgaaa aaccttcaat 4440 ggaggcacaa cgtttattac acgctttacc aacaggtaat gattcaggtg gtgataagct 4500 cttaccaaaa acactttcta aagaaattgt ttcagaacca tttgctaaaa accaattacg 4560 tgaaaaagct cgtctaacta acattaaagg tttagagatt ccaagagttt catatacttt 4620 agacgatgat gacttcatta cagatgtaga aacagcaaaa gaattaaaat taaaaggtga 4680 tacagttaaa ttcactacta ataaattcaa agtatttgct gcaatttcag atactgtaat 4740 tcatggatca gatgtagatt tagtaaactg ggttgaaaac gcactacaat caggtctagc 4800 agctaaagaa cgtaaagatg ccttagcagt aagtcctaaa tctggattag atcacatgtc 4860 attttacaat ggatctgtta aagaagttga gggagcagac atgtatgatg ctattattaa 4920 cgctttagca gatttacatg aagattaccg tgataacgca acaatttata tgcgatatgc 4980 ggattatgtc aaaattatta gtgttctttc aaatggaaca acaaatttct ttgacacacc 5040 agcagaaaaa gtatttggca aaccagtagt atttacagat gcagcagtta aacctattgt 5100 gggagatttc aattattttg gaattaacta tgatggaaca acttatgaca ctgataaaga 5160 tgttaaaaaa ggcgaatatt tgtttgtatt aactgcatgg tatgatcagc aacgtacatt 5220 agacagtgca ttcagaattg caaaagcaaa agaaaataca ggttcattac ccagctaagc 5280 cccaaaaggt taatgtaaca gctaaggcta aatcagctgt aatatcagcc gaataggggt 5340 gatgaaatga gtttagaaga aattaaattg tggttgagaa ttgactataa tttcgaaaat 5400 gatttaattg aaggtctcat tcaatcggct aagtctgaat tactattaag tggggttcca 5460 gattatgaca aagatgactt ggaatacccg cttttttgta cagcgattag atatatcatt 5520 gcaagagatt atgaaagtcg tgggtactca aatgaccaat ctagaagcaa ggtttttaat 5580 gaaaagggat tgcaaaaaat gattctgaaa ttaaaaaagt ggtaggtgat ttttaaatgg 5640 aatttaatga atttaaagat cgcgcatatt tttttcaata tgtaaataaa gggccgtatc 5700 cagatgaaga ggaaaaaatg aagttgtata gttgcttttg taaaatatat aatccttcta 5760 tgaaagatag agaaatttta aaagcgactg aatcaaagtc aggactaacc ataattatga 5820 ggtcttctaa aattgaatat ctaccacaaa caaatcactt agttaaaatt gacagaggct 5880 tatattccga taaattattc aacattaaag aaataagaat tgatacacca gatattggct 5940 ataatacagt ggttttatca gaaaaatgag tgtagaaatt aaagggatac ctgaagtgtt 6000 gaagaaatta gaatcggtat acggtaaaca atcaatgcaa gctaagagtg atagagcttt 6060 aaatgaagca tctgaatttt ttataaaggc tttaaagaaa gaattcgaga gttttaaaga 6120 tacgggtgct agcatagaag aaatgactaa atctaagcct tatacaaaag taggaagtca 6180 agaaagagct gttttaattg aatgggtagg ccctatgaat cgcaaaaaca ttattcactt 6240 gaatgaacat ggttatacaa gagatggaaa aaaatataca ccaagaggtt ttggagttat 6300 tgcaaaaaca ttagctgcta atgaacggaa gtatagagaa attataaaaa aggagttggc 6360 cagataaatg aatatattaa acaccataaa agaaatttta ttatctgatg cagagctcca 6420 aacatatata aattctagaa tatactatta taaagtcact gaaaatgctg aaacttccaa 6480 accttttgtt gttattacac ctatttatga tttaccttca gacttcatgt ctgataaata 6540 tcttagtgaa gaatacttaa ttcaaataga tgtagaatct tcaaataatc agaaaacaat 6600 tgatataaca aaacgaataa gatatctgtt atatcaacaa aatttaattc aagcatctag 6660 tcagttagat gcttattttg aagaaactaa acgttatgtg atgtcgagac gttatcaagg 6720 cataccaaaa aatatatatt ataaaaatca gcgcatcgaa taggtgtgct ttttaatttt 6780 taaggaggaa ataagcaatg gcagaaggac aaggttctta taaagtaggt tttaaaagat 6840 tatacgttgg agtttttaac ccagaagcaa caaaagtagt taaacgcatg acatgggaag 6900 atgaaaaagg tggtacagtt gatctaaata tcacaggttt agcaccagat ttagtagata 6960 tgtttgcatc taacaaacgt gtttggatga aaaaacaagg tactaatgaa gttaagtctg 7020 acatgagtat ttttaatatt ccaagtgaag atctaaatac agttattggt cgttctaaag 7080 ataaaaatgg tacatcttgg gtaggagaga atacaagagc accatacgta acagttattg 7140 gagaatctga agatggttta acaggtcaac cagtgtacgt tgcgctactt aaaggtactt 7200 ttagcttgga ttcaattgaa tttaaaacac gaggagaaaa agcagaagca ccagagccaa 7260 caaaattaac tggtgactgg atgaacagaa aagttgatgt tgatggtact ccacaaggta 7320 ttgtatacgg gtatcatgaa ggtaaagaag gagaagcaga attcttcaaa aaagtattcg 7380 ttggatacac ggacagtgaa gatcattcag aggattctgc aagttcgtta cccagctaac 7440 ccccaaaatg ttgaagtagc agttaattca aaatctgcaa cagtttcagc agaatagggg 7500 ctttcaaaat aaatcaaagg agaataattt atgactaaaa ctttaaaggt ttataaagga 7560 gacgacgtcg tagcttctga acaaggtgaa ggcaaagtgt cagtaacttt atctaattta 7620 gaagcggata caacttatcc aaaaggtact taccaagtgg catgggaaga aaatggtaaa 7680 gaatctagta aagttgatgt acctcaattc aaaaccaatc caattctagt ctcaggcgta 7740 tcatttacac ccgaaactaa atcaatcacg gtaaatgctg atgacaatgt tgaaccaaac 7800 attgcaccaa gtacagcaac gaataaaacg ttgaaatata caagtgaaca tccagagttt 7860 gttactgttg atgagagaac aggagcaatt cacggtgtag ctgagggaac ttcagttatc 7920 actgctacgt ctactgacgg aagtgacaag tctggacaaa ttacagtaac agtaacaaat 7980 ggataattat ttgagacgca gaatatctgc gtctttttta tttgaataaa aggagctaat 8040 acaatgatta aatttgaaat taaagaccgt aaaacaggaa aaacagagag ctatacaaaa 8100 gaagatgtga caatgggcga agcagaaaaa tgctatgagt atttagaatt agtaaatcaa 8160 gagaataaaa aagaagtacc taacgcaaca aaaatgagac aaaaagagcg acagttatta 8220 gtagatttat ttaaagatga aggattgact gaagaagatg ttttgaacaa gatgagcact 8280 aaaacttata caaaagcctt gaaagatata tttcgagaaa tcaatggtga agatgaagaa 8340 gattcagaaa ctgaaccaga agagatggga aagacagaag aacaatctca ataaaagata 8400 ttttatcgaa cattaagaaa atacaacgtt tctgtatgga gcagtatggg tggacattaa 8460 ctgaagtcag aaaacagccg tatgtaaaac ttttagaaat acttaatgaa gagaataaag 8520 aagagactga agaaaaacaa agtgaacaaa aagtcattac aggtacggat ttaagaaaac 8580 tttttggaag ctagaaagga ggttaatatg aatgaaaaag tagaaggcat gaccttggag 8640 ctgaaattag accatttagg tgtccaagaa ggcatgaagg gtttaaagcg acaattaggt 8700 gttgttaata gtgaaatgaa agctaatctg tcatcatttg ataagtctga aaaatcaatg 8760 gaaaagtatc aggcgagaat taaggggtta aatgataagc ttaaagttca aaaaaagatg 8820 tattctcaag tagaagatga gcttaaacaa gttaacgcta attatcaaaa agctaaatct 8880 agtgtaaaag atgttgagaa agcatattta aagctagtag aagctaataa aaaagaaaaa 8940 ttagctcttg ataaatctaa agaagcctta aaatcttcga atacagaact taaaaaagct 9000 gaaaatcaat ataaacgtac aaatcaacgt aaacaagatg catatcaaaa acttaaacag 9060 ttgagagatg cagaacaaaa gcttaagaat agtaaccaag ctactactgc acaactaaaa 9120 agagcaagtg acgcagtaca gaagcagtcc gctaagcata aagcacttgt tgaacaatat 9180 aaacaagaag gcaatcaagt tcaaaaacta aaagtacaaa atgataatct ttcaaaatca 9240 aacgaaaaaa tagaaaattc ttacgctaaa actaatacta aattaaagca aacagaaaaa 9300 gaatttaatg atttaaataa tactattaag aatcatagcg ctaatgtcgc aaaagctgaa 9360 acagctgtta acaaagaaaa agctgcttta aataatttag agcgttcaat agataaagct 9420 tcatccgaaa tgaagacttt taacaaagaa caaatgatag ctcaaagtca tttcggcaaa 9480 cttgctagtc aagcggatgt catgtcaaag aaatttagtt ctattggaga taaaatgact 9540 tccctaggac gtacgatgac gatgggcgta tctacaccga ttactttagg gttaggtgca 9600 gcattaaaaa caagtgcaga cttcgaaggg caaatgtctc gagttggagc gattgcacaa 9660 gcaagcagta aagacttaaa aagcatgtct aatcaagcgg ttgacttagg cgctaaaaca 9720 agtaaaagtg ctaacgaagt tgctaaaggt atggaagaat tggcagcttt aggctttaat 9780 gccaaacaaa caatggaggc tatgccgggt gttatcagtg cagcagaagc aagcggtgca 9840 gaaatggcta caactgcaac tgtaatggca tcagcaatta attctttcgg tttaaaagca 9900 tctgatgcaa accatgttgc tgatttactt gcgagatcag ctaatgatag tgctgcagat 9960 attcaataca tgggagatgc attaaaatat gcaggtactc cagcaaaagc attaggagtt 10020 tcaatagagg acacttctgc agcaattgaa gttttatcta actcagggtt agaggggtct 10080 caagcaggta ctgcattaag agcttcgttt attaggctag ctaatccaag taaaagtaca 10140 gctaaggaaa tgaaaaaatt aggtattcat ttgtctgatg ctaaaggtca atttgttggc 10200 atgggtgaat tgattagaca gttccaagac aacatgaaag gcatgacgag agaacaaaaa 10260 ctagcaacag tggctacaat agttggcact gaagcagcaa gtggattttt agccttgatt 10320 gaagcgggtc cagataaaat taatagctat agcaaatcat tgaagaactc taatggtgaa 10380 agtaaaaaag cagctgattt gatgaaagac aacctcaaag gtgctctgga acaattaggt 10440 ggcgcttttg aatcgttagc aattgaagtt ggtaaagatt taacgcctat gattagagca 10500 ggtgcggaag gattaacaaa attagttgat ggatttacac atcttcctgg ttggtttaga 10560 aaggcttcgg taggtttagc gatttttggt gcatctattg gccctgctgt tcttgctggt 10620 ggcttattaa tacgtgcagt tggaagcgcg gctaaaggct atgcatcatt aaatagacgc 10680 attgctgaaa atacaatact gtctaatacc aattcaaaag caatgaaatc tttaggtctt 10740 caaaccttat ttcttggttc tacaacagga aaaacgtcaa aaggctttaa aggattagcc 10800 ggagctatgt tgtttaattt aaaacctata aatgttttga aaaattctgc aaagctagca 10860 attttaccgt tcaaactttt gaaaaacggt ttaggattag ccgcaaaatc cttatttgca 10920 gtaagtggag gcgcaagatt tgctggtgta gccttaaagt ttttaacagg acctataggt 10980 gctacaataa ctgctattac aattgcatat aaagttttta aaaccgcata tgatcgtgtg 11040 gaatggttca gaaacggtat taacggttta ggagaaacta taaagttttt tggtggcaaa 11100 attattggcg gtgctgttag gaagctagga gagtttaaaa attatcttgg aagtataggc 11160 aaaagcttca aagaaaagtt ttcaaaggat atgaaagatg gttataaatc tttgagtgac 11220 gatgaccttc tgaaagtagg agtcaacaag tttaaaggat ttatgcaaac catgggcaca 11280 gcttctaaaa aagcatctga tactgtaaaa gtgttgggga aaggtgtttc aaaagaaaca 11340 gaaaaagctt tagaaaaata cgtacactat tctgaagaga acaacagaat catggaaaaa 11400 gtacgtttaa actcgggtca aataacagaa gacaaagcaa aaaaactttt gaaaattgaa 11460 gcggatttat ctaataacct tatagctgaa atagaaaaaa gaaataaaaa ggaactcgaa 11520 aaaactcaag aacttattga taagtatagt gcgttcgatg aacaagaaaa gcaaaacatt 11580 ttaactagaa ctaaagaaaa aaatgacttg cgaattaaaa aagagcaaga actcaatcag 11640 aaaatcaaag aattgaaaga aaaagcttta agtgatggtc agatttcaga aaatgaaaga 11700 aaagaaattg aaaagcttga aaatcaaaga cgtgacatca ctgttaaaga attgagtaag 11760 actgaaaaag agcaagagcg tattttagta agaatgcaaa gaaacagaaa tgcttattca 11820 atagacgaag cgagcaaagc aattaaagaa gcagaaaaag caagaaaagc aagaaaaaaa 11880 gaagtggaca agcaatatga agatgatgtc attgctataa aaaataacgt caacctttct 11940 aagtctgaaa aagataaatt attagctatt gctgatcaaa gacataagga tgaagtaaga 12000 aaggcaaaat ctaaaaaaga tgctgtagta gacgttgtta aaaagcaaaa taaagatatt 12060 gataaagaga tggatttatc cagtggtcgt gtatataaaa atactgaaaa gtggtggaat 12120 ggccttaaaa gttggtggtc taacttcaga gaagaccaaa agaagaaaag tgataagtac 12180 gctaaagaac aagaagaaac agctcgtaga aacagagaaa atataaagaa atggtttgga 12240 aatgcttggg acggcgtaaa aactaaaact ggcgaagctt ttagtaaaat gggcagaaat 12300 gctaatcatt ttggcggcga aatgaaaaaa atgtggagtg gaatcaaagg aattccaagc 12360 aaattaagtt caggttggag ctcagccaaa agttctgtag gatatcacac taaggctata 12420 gctaatagta ctggtaaatg gtttggaaaa gcttggcaat ctgttaaatc gactacagga 12480 agtatttaca atcaaactaa gcaaaagtat tcagatgcct cagataaagc ttgggcgcat 12540 tcaaaatcta tttggaaagg gacatcaaaa tggtttagca atgcatataa aagtgcaaag 12600 ggctggctaa cggatatggc taataaatcg cgctcgaaat gggataatat ttctagtaca 12660 gcatggtcga atgcaaaatc cgtttggaaa ggaacatcga aatggtttag taactcatac 12720 aaatctttaa aaggttggac tggagatatg tattcaagag cccacgatcg ttttgatgca 12780 atttcaagtt cggcatggtc taacgctaaa tcagtattta atggttttag aaaatggcta 12840 tcaagaacat atgaatggat tagagatatt ggtaaagaca tgggaagagc tgcggctgat 12900 ttaggtaaaa atgttgctaa taaagctatt ggcggtttaa atagcatgat tggcggtatt 12960 aataaaatat ctaaagccat tactgataaa aatctcatca agccaatacc tacattgtct 13020 actggtactt tagcaggaaa gggtgtagct accgataatt cgggagcatt aacgcaaccg 13080 acatttgctg tattaaatga tagaggttct ggaaacgccc caggtggtgg agttcaagaa 13140 gtaattcaca gggctgacgg aacattccat gcaccccaag gacgagatgt ggttgttcca 13200 ctaggagttg gagatagtgt aataaatgcc aatgacactc tgaagttaca gcggatgggt 13260 gttttgccaa aattccatgg tggtacgaaa aagaaagatt ggctagacca acttaaaggt 13320 aatataggta aaaaagcagg agaatttgga gctacagcta aaaacacagc gcataatatc 13380 aaaaaaggtg cagaagaaat ggttgaagca gcaggcgata aaatcaaaga tggtgcatct 13440 tggttaggcg ataaaatcgg cgatgtgtgg gattacgtac aacatccagg gaaactagta 13500 aataaagtaa tgtcaggttt aaatattaat tttggaggcg gactaacgct acagtaaaaa 13560 ttgctaaagg cgcgtactca ttgctcaaaa agaaattaat agacaaagta aaatcgtggt 13620 ttgaagattt tggtggtgga ggcgatggaa gctatctatt tgaatatcca atctggcaaa 13680 gatttggacg ctacacaggt ggacttaact ttaatgacgg tcgtcactat ggtatagact 13740 ttggtatgcc tactggaaca aacgtttatg ccgttaaagg tggtatagca gataaggtat 13800 ggactgatta cggtggcggt aattctatac aaattaagac cggtgctaac gaatggaact 13860 ggtatatgca tttatctaag caattagcaa gacaaggcca acgtattaaa gctggtcaac 13920 tgatagggaa atcaggtgct acaggtaatt tcgttagagg agcacactta catttccaat 13980 tgatgcaagg gtcacatcca gggaatgata cagctaaaga tccagaaaaa tggttgaagt 14040 cacttaaagg tagtggcgtt cgaagtggtt caggtgttaa taaggctgca tctgcttggg 14100 caggcgatat acgtcgtgca gcaaaacgaa tgggtgttaa tgttacttcg ggtgatgtag 14160 gaaatatcat tagcttgatt caacacgaat caggaggaaa tgcaggtata actcaatcta 14220 gttcgcttag agacatcaac gttttacagg gcaatccagc aaaaggattg cttcaatata 14280 tcccacaaac atttagacat tatgctgtta gaggtcacaa caatatatat agtggttacg 14340 atcagttatt agcgttcttt aacaacagat attggcgctc acagtttaac ccaagaggtg 14400 gttggtctcc aagtggtcca agaagatatg cgaatggtgg tttgattaca aagcatcaac 14460 ttgctgaagt gggtgaagga gataaacagg agatggttat ccctttaact agacgtaaac 14520 gagcaattca attaactgaa caggttatgc gcatcatcgg tatggatggc aagccaaata 14580 acatcactgt aaataatgat acttctacag ttgaaaaatt gttgaaacaa attgttatgt 14640 taagtgataa aggaaataaa ttaacagatg cattgattca aactgtttct tctcaggata 14700 ataacttagg ttctaatgat gcaattagag gtttagaaaa aatattgtca aaacaaagtg 14760 ggcatagagc aaatgcaaat aattatatgg gaggtttgac taattaatgc aatcttttgt 14820 aaaaatcata gatggttaca aggaagaagt aataacagat tttaatcagc ttatattttt 14880 agatgcaagg gctgaaagtc caaacaccaa tgataacagt gtaactatta acggagtaga 14940 tggtatttta ccgggcgcaa ttagttttgc gcctttttca ttagtattaa ggtttggcta 15000 tgatggtata gatgttatag atttaaattt atttgagcat tggtttagat ctgtgtttaa 15060 tcgcagacat ccttattatg ttattacttc tcaaatgcct ggtgttaaat atgcagtgaa 15120 tacagctaat gttacatcta atttaaaaga tggttcttca actgaaattg aagtaagttt 15180 aaatgtttat aaagggtatt ctgaatcagt taattggacc gatagcgagt tcttattcga 15240 ctctaattgg atgtttgaaa atggaattcc tcttgatttc acacctaaat atactcatac 15300 atcaaatcaa tttactattt ggaacggttc tactgatacg ataaatccac gattcaagca 15360 cgatttgaaa atattaatta atttaaatgc gagtggagga tttgaactgg ttaactatac 15420 aacaggtgat atttttaagt acaacaaaag tatagataaa aacactgatt ttgttttaga 15480 tggtgtgtat gcatatcgag atataaatag agtgggaatt gatacaaata gaggcattat 15540 aacattagcg ccaggtaaaa atgaatttaa gattaaagga gacatcagtg atattaaaac 15600 tacatttaag tttcctttta tttataggta ggtgatttaa tggattatca tgatcattta 15660 tcagtaatgg attttaatga attgatttgt gaaaatttac tagatgtaga ttatggttct 15720 tttaaagaat attatgaact gaatgaagct aggtacatca cttttacagt ttatagaact 15780 actcataata gttttgtttt cgatttacta atttgtgaaa acttcataat ttatcatggt 15840 gaaaaataca caattaagca gacagcgcca aaggttgaag gtgataaagt ttttattgaa 15900 gttacggcat atcacataat gtatgaattt caaaatcact cagtggaatc aaataagctt 15960 gatgacgaca gtagcgaaac tggtaaaacg ccagaatact ctttagatga gtacttaaga 16020 tatggatttg caaatcaaaa aacttcggtc aaaatgacct ataaaataat tggaaatttt 16080 aagcgaaaag taccgattga cgaattaggt aacaaaaacg gcttagaata ctgtaaagaa 16140 gcggtagacc tatttggctg tataatttac ccaaatgata cggagatatg tttttattct 16200 cctgaaacat tttatcaaag aagcgagaaa gtgattcgat atcaatataa tactgatact 16260 gtatctgcaa ctgtcagtac attggaatta agaacagcta taaaagtttt tggaaaaaag 16320 tatacagctg aggaaaagaa aaattataat cctattagaa caactgacat taaatattca 16380 aatggtttta taaaagaagg tacttatcgt accgcaacaa ttgggtctaa agctactatt 16440 aactttgatt gcaagtatgg taatgaaaca gttagattta caataaaaaa gggctctcaa 16500 ggtggaatat ataagttgat tttagacggc aagcaaatta agcaaatttc ttgttttgct 16560 aagtcggttc agtctgaaac aatagattta ataaaaaata ttgataaagg caagcacgtt 16620 ttagaaatga tatttttagg agaagacccc aaaaatagaa ttgatatatc ttcaaataaa 16680 aaagctaagc cttgtatgta tgttggaact gaaaaatcaa cagtcttaaa tttaattgct 16740 gacaactcag gtcgcaatca atacaaagca attgttgact acgtcgcaga tagtgcaaag 16800 cagtttggga ttcgatatgc taatacgcaa acaaatgaag atatcgaaac acaggataag 16860 ctgttagaat ttgcaaaaaa gcaaataaat gatactccta agactgaatt agatgttaat 16920 tatataggtt atgaaaaaat agagccaaga gatagcgtat tctttgttca tgaattaatg 16980 ggatataaca ctgaattaaa ggttgttaaa cttgataggt cacatccatt tgtaaacgca 17040 atagatgaag tgtctttcag caatgaaata aaggatatgg tacaaattca acaagcgctt 17100 aacagacgag ttattgcaca agataataga tataactatc aagcaaatcg tataaatcat 17160 ttatacacta gtactttgaa ttctcctttc gagacaatgg atatagggag tgtattaata 17220 taatggcaac agaagaagtt aaaatcaaag cgctacttga aaacgataaa cagtactttc 17280 cagctacaca ttggaaagct ataaatggga taccttatgc aggcagtagt gatattgatg 17340 gattgcctca agacggtatc atttcggtag atgataaaaa taaattagat aatttaaaaa 17400 taggcgaagc aggaattatt caaaatagca ttgtacagaa atccccaaac ggtaaattgt 17460 ggaaaataac agttgacgat agtgggaaac ttggtacagt gctattttat tagaaaggaa 17520 ggtgcattat ggaaaatttg tatttaataa aggatttggg agctttagca ggtcgagatt 17580 atagagctaa ggaaatacaa aacttacaaa gaatagagca atttgcgctt ggcttgacaa 17640 cagagtttaa gttgcatcag aaagctaaaa caattcaaca cttcgctgag caaatttatt 17700 ataatggtag atcgcaagca gcagtaaaca aatctttaca aagtcaaatt aacgcacttg 17760 ttgtggcacc acgtaataac agtgctaatg agattgttca agctcgagtt aatgtaaacg 17820 gcgaaacctt tgacacatta aaagaacatt tagacgattg ggaaacccaa actcaaatta 17880 ataaagagga aactataaga gaattaaata agaccaaaca agaaattctt gatatcgagt 17940 atcgttttga acctgataag caagaatttt tatttgtgac agaacttgca cctcttacaa 18000 atgcagtaat gcaatccttc tggtttgata atagaacagg catagtatac atgacacaag 18060 ctagaaataa tggctatatg ctaagtcgtc taagacctaa tggtcaattt atagacagct 18120 cattgattgt aggtgggggt catggtacac ataacggtta tagatatatt gatgatgagt 18180 tatggattta tagttttatc ttaaatggta ataatgagaa tacattagtt cgtttcaagt 18240 atacgcctaa tgtggaaatt agctatggca agtatggtat gcaagatgta tttacaggac 18300 acccagaaaa accctacatc acccctgtca taaatgaaaa agaaaataaa attctataca 18360 gaattgagag acctagaagt cactgggaac ttgaaaactc aatgaattat atagagataa 18420 gaagtttaga cgatgttgat aaaaatattg ataaagtttt gcataaaatc agtatcccta 18480 tgagactaac aaacgaaacc caaccaatgc agggtgtgac ttttgatgaa aaatacttgt 18540 attggtatac aggagacagt aatccaaata atagaaacta tttaacggct ttcgatttag 18600 aaacaggaga agaagcgtat caggttaatg ctgactatgg tggaacacta gattcatttc 18660 ctggcgaatt tgcggaagca gaaggtttgc aaatatacta tgacaaagat agtggtaaaa 18720 aagctttgat gctaggtgtt actgtcggtg gtgatggaaa tagaacacat cgtattttca 18780 tgattgggca aagaggtatt ttagaaatac ttcactcaag aggcgttcct tttatcatga 18840 gtgacacagg tggtagagtt aaacctttac caatgaggcc tgataaactt aagaatcttg 18900 ggatgttaac agagccaggt ctttactatt tatacactga tcatacagtt caaatcgatg 18960 atttcccatt accaagagaa tggcgtgatg caggttggtt cttggaagtt aagccaccac 19020 aaactggcgg tgatgtaatt cagatattga cgcgtaatag ttatgcaagg aatatgatga 19080 cttttgaaag ggtgctttct ggaagaactg gagacatttc ggactggaat tatgtgccta 19140 aaaatagtgg taaatgggag agagtacctt cattcatcac aaaaatgtca gatattaaca 19200 tagtaggcat gtcgttttat ttaactacgg atgatacaaa acgttttaca gattttccaa 19260 ctgaacgtaa aggggtagct ggttggaact tatatgtaga agcttcaaac acaggtggct 19320 ttgttcatag gctagttcgt aatagtgtta cagcatctgc tgagatacta ttgaaaaatt 19380 atgatagtaa aacaagttca gggccatgga ctttacacga agggagaatt ataagttaat 19440 gagtaattta gagaaatctg tagctataaa tttagaaaac acagcgcatt atgaaaatat 19500 ttcaaatcta gatataactt ttagaacagg agagagtgat tcttctgttc ttctttttaa 19560 tatcactaaa aataatcaac cgttattatt gagtgaagaa aatatcaaag cacgaatagc 19620 gattcgaggt aaaggagtca tggtagttgc tccactagaa atattagatc catttaaagg 19680 tattttaaaa tttcaattac ctaatgatgt aattaaacga gatggaagtt atcaagctca 19740 agtttcggtt gcagaattag gtaattcaga cgtggtagtt gtcgagagaa ctatcacatt 19800 taacgttgaa aaaagtttgt ttagcatgat tccatctgaa acaaaattac actatattgt 19860 tgaatttcag gaattagaaa aaactattat ggatcgtgcg aaagcaatgg acgaggctat 19920 aaaaaatggt gaagattatg cgagtctgat tgaaaaagct aaagaaaaag gtctatcaga 19980 tattcaaata gcaaaatctt caagtataga tgaattaaag caacttgcta atagccatat 20040 atctgatttg gaaaataaag cgcaagcata ttcaagaaca ttcgatgagc aaaagcgata 20100 tatggatgag aaacatgaag ccttcaagca gtcagtgaat agtggtggtt tagtcacaag 20160 tggttctact tcaaattggc aaaaagctaa gattactaaa gatgatggta agataatgca 20220 gattactgga tttgatttta ataatccaga acaaagaata ggtgattcaa cccaatttat 20280 ttatgtttcg caagctataa attatccaag aggtgttagt actaacggta ctgtcgaata 20340 tttagtagta acttcagatt acaagcgtat gacttatcga ccgaacggta caaataaagt 20400 gtttgttaaa agaaaagaag cgggttcatg gtctgagtgg tcagaattag ctattaatga 20460 ttacaataca ccttttgaaa ctgttcaaag tgcccaatca aaagctaata tggccgaaag 20520 taacgctaaa ttatacgcag atgacaagtt taataaaagg tattcggtta tttttgatgg 20580 aacagcaaat ggtgtgggct ctacattgta cttaaatgag agtttagacc aatttatttt 20640 attaattttt tatgggactt ttccaggtgg tgactttaca gagtttggca gtccttttgg 20700 aggaggaaag atttcattga atccctcaaa tcttccagat ggtgatggaa atggtggagg 20760 tgtttatgag tttggattaa ctaaatctag tcgtacatct ttaactatat caaacgatgt 20820 ctatttcgac ttaggaagtc aaagaggctc tggtgcgaac gcaaatagag ggacaattaa 20880 caaaattata ggagtgagaa aataatgcaa atattagtta acaagcgtaa tgagataatt 20940 tcatacgcta tcattggtgg ctttgaagaa ggtattgata ttgaaaattt accagaaaat 21000 ttctctcaag tttttagacc taaagccttt aaatattcaa atggggaaat agtttttaac 21060 gaagattatt cagaagaaaa agatgacttg catcaacaga ttgacagtga agaacaaaac 21120 acagtcgctt ctgatgacat cttacgaaaa atggttgcta gtatgcagaa acaagttgtt 21180 caaagtacaa agttatcgat gcaagttaat aagcaaaatg cactaatggc aaaacaactt 21240 gtgacactta ataaaaaatt agaagaggtt aaaggagaga ctgaaaatgc ttaaattaat 21300 ttcaccaaca ttcgaagata ttaaaacatg gtatcaattg aaagaatata gtaaagaaga 21360 tatagcgtgg tatgtagata tggaagttat agataaagag gaatatgcaa ttattacagg 21420 agaaaagtat ccagaaaatc tagagtcata ggttataatc ttatggcttt ttaatttgaa 21480 taaagtgggt ggtgtaatgt ttggatttac caaacgacac gaacaagatt ggcgtttaac 21540 gcgattagaa gaaaatgata agactatgtt tgaaaaattc gacagaatag aagacagtct 21600 gagaacgcaa gaaaaaattt atgacaagtt agatagaaat ttcgaagaac taaggcgtga 21660 caaagaagaa gatgaaaaaa ataaagagaa aaatgctaaa aatattagag acatcaagat 21720 gtggattcta ggattaatag ggacgattct aagtacattt gttatagcct tgttaaaaac 21780 tatttttggc atttaaagga ggtgattacc atgcttaagg gaattttagg atatagcttt 21840 tggtcgtgtt tctggtttag taagtgtaag taatagttaa gagtcagtgc ttcggcactg 21900 gctttttatt ttggaaaaaa ggagcaaaca aatggatgca aaagtaataa caagatacat 21960 cgtattgatc ttagcattag taaatcaatt cttagcgaac aaaggtatta gcccgattcc 22020 agtagacgat gagaatatat catcaataat acttactgtt gttgctttat atactacgta 22080 taaagacaat ccaacatctc aagaaggtaa atgggcaaat caaaagctaa agaaatataa 22140 agctgaaaac aagtatagaa aagcaacagg gcaagcgcca attaaagaag taatgacacc 22200 tacgaatatg aacgacacaa atgatttagg gtaggtgttg accaatgttg ataacaaaaa 22260 accaagcaga aaaatggttt gataattcat tagggaagca gttcaatcct gatttgtttt 22320 atggatttca gtgttacgat tacgcaaata tgttttttat gatagcaaca ggcgaaaggt 22380 tacaaggttt atacgcttat aatattccat ttgataataa agcaaggatt gaaaaatacg 22440 ggcaaataat taaaaactat gatagctttt taccgcaaaa gttggacatt gtcgttttcc 22500 cgtcaaagta tggtggcgga gctggacatg ttgaaattgt tgagagcgct aatctaaaca 22560 ctttcacatc gtttggccaa aattggaatg gtaaaggttg gacaaatggc gttgcgcaac 22620 ctggttgggg tcccgaaacc gttacaagac atgttcatta ttacgatgac ccaatgtatt 22680 ttattagatt aaatttccca gataaagtaa gtgttggaga taaagctaaa agcgttatta 22740 agcaagcaac tgccaaaaag caagcagtaa ttaaacctaa aaaaattatg cttgtagccg 22800 gtcatggtta taacgatcct ggagcagtag gaaacggaac aaacgaacgc gattttatac 22860 gtaaatatat aacgccaaat atcgctaagt atttaagaca tgccggtcat gaagtcgcat 22920 tatatggtgg ctcaagtcaa tcacaagaca tgtatcaaga tacagcatac ggtgttaatg 22980 taggtaataa aaaagattat ggcttatatt gggttaaatc acaggggtat gacattgttc 23040 tagaaataca tttagacgca gcaggagaaa gcgcaagtgg tgggcatgtt attatctcaa 23100 gtcaattcaa tgcagatact attgataaaa gtatacaaga tgttattaaa aataacttag 23160 gacaaataag aggtgtaaca cctcgtaacg atttactaaa tgttaacgta tcagcagaaa 23220 taaatataaa ttatcgctta tctgaattag gttttatcac taataaaaat gatatggatt 23280 ggattaagaa aaactatgac ttgtattcta aattaatagc cggtgcgatt catggtaagc 23340 ctatcggtgg tgtgatatct agtgaggtta aaacaccagt taaaaacgaa aagaatccgc 23400 cagtgccagc aggttataca cccgataaaa ataatgtacc gtataaaaaa gaaactggtt 23460 attacacagt tgccaatgtt aaaggtaata acgtaaggga cggctattca actaattcaa 23520 gaattactgg tgtattacct aataacgcaa caatcaaata tgacggcgca tattgtatca 23580 atggctatag atggattact tatattgcta atagtggaca acgtcgttat attgctacag 23640 gagaggtaga caaggcaggt aatagaataa gcagttttgg taagtttagt gcagtttgat 23700 aattgtatat gatgaatctt aggcaggtac ttcggtactt gcctattatt taaaattaat 23760 aaacagttaa tttttacatg aatatattaa attttaaaaa aacaaacgtt tttagtatat 23820 aaattatttt gtgttcgtat tgtgtgctat gattaaaaag ttgttatggt caactatatc 23880 gtggttttat gtttattatc aatcaaaata taaattattt ataatttgtt tggtaatgaa 23940 cgggtttttt tcgaaataat agtaaaaaaa cacatttgta gatattttaa actcggtaaa 24000 tcttttaata aatatttaat tttattaaaa gttaaaaagg tttaatataa aaatgtaata 24060 aaatttataa agaaaggaaa tgatttttat ggtcaaaaaa agactattag ctgcaacatt 24120 gtcgttagga ataatcactc ctattgctac ttcgtttcat gaatctaaag ctgataacaa 24180 tattgagaat attggtgatg gcgctgaggt agtcaaaaga acagaagata caagtagcga 24240 taagtggggg gtcacacaaa atattcagtt tgattttgtt aaagataaaa agtataacaa 24300 agacgctttg attttaaaaa tgcaaggttt tatcaattca aagactactt attacaatta 24360 caaaaacaca gatcatataa aagcaatgag gtggcctttc caatacaata ttggtctcaa 24420 aacaaatgac cccaatgtag atttaataaa ttatctacct aaaaataaaa tagattcagt 24480 aaatgttagt caaacattag gttataacat aggtggtaat tttaatagtg gtccatcaac 24540 aggaggtaat ggttcattta attattcaaa aacaattagt tataataaaa taaaaagtag 24600 gtgataagat gactcaattt ctaggggcgc ttcttcttac aggagtttta ggttacatac 24660 catataaata tctaacaatg ataggtttag ttagtgaaaa aaacaaggtt atcaatactc 24720 ctgtattatt gattttttct attgaaacat gtttgatatg gttttatagt tttataattt 24780 ttaataatgt tgatttaaaa aatttgaatt taattcagtt gcttacaggt ctaaaagcaa 24840 atattttgtt tctatttatt tttgttttaa cagtgtttgt atttaatcct ttaattgtta 24900 aatttattat ctggttaatt aatataacca gaaagtttat gaaattggat tgtataagct 24960 tattagacaa aagagacaag ttgtttaata acaacggtaa accagtattt atagttataa 25020 aagactttga aaacagaatc attgaagagg gtgaacttaa aacctataat tcagctggta 25080 gcgatttcga tttactagaa gttgagcgac aagatttcaa agtatctgat ttaccgtcaa 25140 acgatgaatt gtatattaaa catacacttg tagaccttaa acaacaaatt aaattggatt 25200 tatatttaat gaatgaatac taatcttttt tcttagcttt ttctgataaa gtgcttttta 25260 atttttcgct ggcgcccggc ttttcaaaac ttttgtttat tgggttacta cgagtagctt 25320 cttgtttttt gtttttatcc gccataaaat tctcaccacc attcaacgtc tacacttgta 25380 ggcgtttttt tatttagtaa agtcataatg aatcttcttt ggttaactta tctccatcta 25440 ttttttgtga aataaattcc aagtatttac gcgcattatg tgacgataaa tctttaggta 25500 actcataagt gaatggttga ttaccactag ttaaaacttc atatactata gtttcttttt 25560 ttattttgca attagttatt ttcattataa acttcctttc aaacactgct gaaatagacg 25620 tcttttatat taaagcgcca cacaggcgct gttaatcaca atacaacttt gcccattact 25680 ttaatattac taaacgaagc gactttgata tcatcatact tcggatttag agataccaaa 25740 ttaatatagt cttcgcatat atctacacgc ttgataagac ttactccatc taatacaacg 25800 agtgcaattg taccatcttt aatagaatct tctttcttaa taaaagcgta tgttccttgt 25860 tttaacatag gttccattga atcaccatta actaaaatac aaaaatcagc atttgatggc 25920 gtttcgtctt ctttaaaaaa tacttcttca tgcaatatgt catcatataa ttcttctcct 25980 atgccagcac cagttgcacc acatgcaata tacgatacta gtttagactc tttatatcca 26040 tctatagaag tgactttatt ctgttcttcc aattgttcat ttgcatagtt aagtacgttt 26100 tcttggcggg gaggtgtgag tttgttgtat atggaagtga tgtcgttatc gtctttgtat 26160 gtagtatttg attcactata caaatcatta atcttcacat tgaagtactc agccaaaatt 26220 ttggcagttg ataatcgagg ttcttccttt tcattttccc attttgatat cttgcctttc 26280 gttaatttca ttaagtcggg atatttatta ttaagatcag ttgctaattg ttccatagtc 26340 atatttttat ttttttctta gcttctttaa accttcacca atacccatac gaaaccctcc 26400 ttatataaga taatttcatt ataaaagttt cgaaaacgaa acgcaaggaa aatattattg 26460 caaaagttgt tgacatcgaa acttttatga tgtattctta aatcaagttg ttacaaacga 26520 aacaaaagga gggggttcaa tgacaactag tgtagcagat aaaccatact taaaaataaa 26580 aagcttgatt gcacttaaag gaactaacca aaaagaagtt gctaaagcaa tcggaatgag 26640 tagaagttta ttgagtataa agataaatcg aattaatggc agagatttta caacttcaga 26700 agctaaaaaa ttagcagatc atttaaatgt taaagttgat gatttttttt aaactttaag 26760 tttcgaaagt gacaactaaa taaaaataag gaggacacta tggaacaaat aacgttaacc 26820 aaagaagagt tgaaagaaat tatagcgaaa gaagttagaa atgctataaa aggcgagaaa 26880 ccaatcagct caggtgcaat tttcagtaaa gtaagaatca ataatgacga tttagaagaa 26940 atcaataaaa aactcaattt cgcaaaagat ttgtcgctag gaagattgag gaagctcaat 27000 catccgattc cgctaaaaaa gtatcagcat ggcttcgaat caattcatca aaaagcttat 27060 gtacaagatg ttcatgacca tattagaaaa ttaacattat caatttttgg agtgacactt 27120 aattcagact tgagtgaaag tgaatacaac ctagcagcaa aaatttatag agatatcaaa 27180 aactattatt tatatatcta tgaaaagaga gtttcagaat taactatcga tgatttcgaa 27240 tgaaggagga actacaaatg aaactactaa gaaggctatt caataaaaaa cacgaaaact 27300 taattgacgt gtggcatgga aatcaatggt taaaagtgaa agaaagcaaa ttaaaaaaat 27360 ataaagtggt ctcggataga gaaggtaaga aatatctaat taaataagcg cacttaatta 27420 gtgcaagtaa tcaagtgcgc tattgcctta caatcctaaa tcttttctgc ttttttcttc 27480 ttcttgtaat cccaataaca cagaagagta aatgctgaaa tagtcacgag caacgctatc 27540 tttagcgaat gcaattacgt catcaccgac ttcttgccat tcgttatgaa tcttatgtct 27600 atctagagct ctaggtaata gcgagattgt aatatcgtga gcaattttct ctaaatccat 27660 aaatttcacc tccttccact gggagataac taaattatat aacaaaacaa cttaaaggag 27720 gaacgacaaa tgcaagctca aaacaaaaaa gtcatctatt actactatga cgaagaaggt 27780 aataggcgac cattagatat tcaaattaat gacggatatg aactgatggt ccgatctcat 27840 ttcatcaaca acaccattga agaaatacca tacgtaaata ataacttata tgccttggtt 27900 gatggttatg aatttaagtt agattgaatt tttgagaaag atattgaaaa gctaatttcc 27960 ccataagatt aagagacata ctggatgttt tgttaacgac tcttttaact tcgttccaag 28020 ttttattgtc tctaatatta tcgagaaatt catggccaga ccaagtgatg tcatcaataa 28080 tccaagaaac gaccctgcct tcgatgaatt tcagatcgca acaaataaat ttagcttctt 28140 ctaattttaa aagtgagtac attactgttt caaaatcata tttatcaaaa ataatattat 28200 cgttgaaatt atgtcgagta agtggttcac ctattttctt attagattct atttctaaga 28260 gcaagagtct aacgcaatcg tgattaagtt tcatcctatc acctccataa caggagtata 28320 gcagaaagga tcataaacat cttaaaagga ggaataacaa atgaacattc aagaagcaac 28380 taagatagct acaaaaaatc ttgtctctat gacacggaaa gattggaaag aaagtcatcg 28440 aactaagata ttaccaacaa atgatagttt tttacaatgc atcatttcaa atagcgatgg 28500 gacaaacctt atcagatatt ggcaaccttc agccgatgac ctcatggcaa atgattggga 28560 agttataaac ccaactagag accaggaatt attgaagcaa ttttagaaat gctatcaatg 28620 atacttttta aattgttttt aaactcattt tcaaagtaaa caacagtctt gtctgaaatt 28680 gttacatgat aaatagtgtt actagcatac acgccgttta ggaacccaga gtttttaagt 28740 ttatttaaat cgtattttac atcttcgaaa tgtagttttt gaaaatactt tgtatgtata 28800 tctttagcac ttccaaaatt attgcaggtt aatttaaccg aacctaactt tacacattct 28860 aaataatctt tgtagagtac ggacaagata tattgttggt ctttagtaag tgtatcaaat 28920 tcatcagata tcaagggcat gttatcacct ccttaggttg ataacaacat tatacacgaa 28980 aggagcataa acaaatgaac acaagatcag aaggattgcg tataggcgtc ccacaagttt 29040 ctagcaaagc tgatgcttct tcatcctatt taacggaaaa ggaacgtaac ttaggagcgg 29100 aaatattaga gcttattaaa aaaagtgatt acagctactt agaaataaac aaagttttct 29160 atgcattaga tagagaactt caatacaggg cgaataataa caaactttaa catttatcta 29220 aaggagtgat agagatgcca aaaatcataa taccaccaac accagaaaac acatatcgag 29280 gcgaagaaaa atttgtgaaa aagttatacg caacacctac acaaatccat caattgtttg 29340 gagtatgtag aagtacagta tacaactggt tgaaatatta ccgtgaagat aatttaggtg 29400 tagaaaattt atacattgat tattcagcaa cgggaacatt gattaatatt tctaaattag 29460 aagagtattt gatcagaaag cataaaaaat ggtattagga ggattatcaa atgagcgaca 29520 catataaaag ctacctatta gcagtgttgt gcttcacggt cttagcgatt gtactcatgc 29580 cgtttctata cttcactaca gcatggtcaa ttgcgggatt cgcaagtatc gcaacattca 29640 tattttataa ggaatacttt tatgaagaat aaagaaactg ctacttgttg gagcaagtaa 29700 cagtgcaaga tgagcaattg tcttaaataa ttatataagg agttattaat atgaccttac 29760 aacaaaaaat actatcacat tttgcaacat atgacaattt caattctgat gatgttgttg 29820 aagtttttgg gatatctaaa acacatgcaa aatccacact ttcaagactt aagaaaaaag 29880 gaaagattga attggaaagt tggggtatct ggcgtgttgt tgaaccgcag ttacatttaa 29940 ctgttgtaga acgtaagaaa gagatattag aagaacaatt cgagttattg gcaagattaa 30000 acgaacaaag tgatgaccct agagaaatag aagaacgcat caagttaatg attcgtttag 30060 ccaaccaatt ttaaggagga gttaatcaat ggcaatatta gaaggtattt ttgaagaatt 30120 aaaactatta aataagaatt tacgtgtgct aaatactgaa ctatcaactg tagattcatc 30180 aattgtacaa gagaaagtta aagaagcacc aatgccaaaa gatgaaacag ctcaactgga 30240 atcagttgaa gaagttaagg aaacttctgc tgatttaact aaagattatg ttttatcagt 30300 aggaaaagag ttccttaaaa aagcagatac ttctgataag aaagaattta gaaataaact 30360 taacgaactt ggtgcggata agctatctac tatcaaagaa gagcattatg aaaaaattgt 30420 tgattttatg aatgcgagaa taaatgcatg aagctagatc actcaaatag agctcatgca 30480 aagcttagtg caagtggagc aaaacaatgg ctaaactgtc caccgagtat taaggcaagt 30540 gaaggtattg cagataaaag ttcagttttt gctgaagaag gtacattcgc tcatgagtta 30600 agtgagttat atttcagtct taaatatgaa ggcctaacac agtttgagtt taataaagct 30660 tttcaaaatt ataagcgaaa tcaatattac agtgaagagt tgcgcgaata tgttgaagag 30720 tacgtagcta atgtagaaga aaaatataac gaagctttga gtagagatga cgatgtaata 30780 gctttatttg aaacaaaatt ggatttaggt aaatacgtcc ctgaatcttt tggtactggt 30840 gatgtcatta tattttcagg tggtgtactt gaaattattg accttaaata cggtaaaggc 30900 attgaagttt cagctataga taatcctcaa cttagattat atggcttggg cgcatatgaa 30960 ctgcttagtt taatgtatga cattcataca gttcgcatga ctatcataca accacgaata 31020 gataactttt ctactgaaga gttaccaata tcaagattac ttcaatgggg aaccgatttt 31080 gttaaaccat tagccagact tgcttataac ggtgaaggtg agtttaaagc aggtagtcat 31140 tgtagattct gtaagataaa gcattcatgt agaacacgtg cagaatacat gcaaaatgtg 31200 cctcaaaagc caccacattt gttgagtgat gaagagattg cagaactttt atataaactg 31260 cctgacatca aaaaatgggc tgatgaagta gaaaaatatg cactagatca agcgaaagaa 31320 aatgataaaa actattctgg ttggaagctt gtagaaggtc gctcgcgaag aatgataact 31380 gatacaaatg caacgcttga aaagttagtt gaagcaggtt ataaacctga agatattaca 31440 gaaaccaagt tacttagcat tacgaattta gaaaaattaa tcggcaaaaa agcattttct 31500 aaaattgcag aaggctttat agaaaagcca caaggtaaat taacacttgc taccgagtct 31560 gataaacgac cagctataaa gcaatctgct gaagatgatt ttgacaaact ataaaaatta 31620 aaaaggacgg tatataaaca tgaaagcaaa agtattaaat aaaactaaag tgattacagg 31680 aaaagtaaga gcatcatatg cacatatttt tgaacctcac agtatgcaag aagggcaaga 31740 agcaaagtat tcaatcagtt taatcattcc taaatcagat acaagtacga taaaagccat 31800 tgaacaagct atagaagctg ctaaagaaga aggaaaagtt agtaagtttg gaggcaaagt 31860 tcctgcaaat ctgaaacttc cattacgtga tggagatact gaaagagaag atgatgtgaa 31920 ttatcaagac gcttatttta ttaacgcatc aagcaaacaa gcacctggta ttattgacca 31980 aaacaaaatt agattaacgg attctggaac tattgtaagt ggtgactata ttagagcttc 32040 aatcaattta tttccattca acacaaatgg taataagggt atcgcagttg gattgaacaa 32100 cattcaactt gtagaaaaag gcgaacctct tggcggtgca agtgcagcag aagatgattt 32160 cgatgaatta gacactgatg atgaggattt cttataagtc aataggtggg gtttttagcc 32220 ccactttaat tttaaagaaa ttgaggtgtc aagaatttga aatttatgaa tatagatatt 32280 gaaacatata gcagtaacga tatttcgaaa tgtggtgtct ataaatacac agaagctgaa 32340 gatttcgaaa tcttaattat agcttattca atagatggtg gaccgattag tgcgattgac 32400 atgactaaag tagataatga gcctttccac gctgattatg agacgtttaa aattgctcta 32460 tttgaccctg ctgtaaaaaa gtatgcattc aatgctaatt tcgaaagaac ttgtcttgct 32520 aaacatttta ataaacagat gccacctgaa gaatggattt gcacaatggt taattcaatg 32580 cgtattggct tacctgcttc gcttgataaa gttggagaag ttttaagact acaaaaccaa 32640 aaagataaag caggtaaaaa tttaattcgt tatttctcta taccttgtaa gccaacaaaa 32700 gttaatggag gaagaacaag aaatttgcct gaacatgatc ttgaaaaatg gcaacaattt 32760 atagattact gtattcgaga tgtagaagta gaaatgacaa ttgctaataa aattaaagac 32820 tttccagtaa ctgtaattga acaagcatat tgggtttttg accaacatat aaacgacaga 32880 ggtattaagc tttctaaatc attgatgtta ggagctaatg tgctcgataa gcagagtaaa 32940 gaagaattgc ttaaacaagc taaacatata acaggtttag aaaatcctaa tagtcctaca 33000 cagttattgg cttggttaaa ggatgaacaa ggattagata tacctaattt acaaaagaaa 33060 acggttcagg attacttaaa agtagcaaca ggaaaagcta aaaaaatgct agaaattaga 33120 ttgcaaatgt ctaaaaccag tgtgaaaaaa tacaacaaaa tgcatgacat gatgtgcagt 33180 gatgaacggg taagaggtct gtttcaattc tacggtgccg gtactggaag atgggcaggt 33240 agaggtgtac aacttcagaa tttaacaaag cattatattt cagatactga attagaaata 33300 gcaagagatc ttattaaaga acaacgtttt gacgatttag atttattact caatgttcat 33360 cctcaagact tattaagtca attagttagg acgacattta ctgctgaaga aggtaatgaa 33420 ctagcagtaa gtgatttttc tgcaatagag gcaagagtca tagcatggta tgcaaaagaa 33480 caatggcgtt tagatgtgtt caacacacac ggaaagatat atgaagcatc ggcttctcaa 33540 atgtttaatg taccggtaga aagcataact aaaggcgacc ctctcagaca aaaaggaaaa 33600 gtgtccgaat tagctttagg ctatcaaggt ggcgctggag ctttaaaagc aatgggtgca 33660 ttggaaatgg gcattgaaga aaacgagtta caaggtttag ttgatagttg gcgtaacgca 33720 aatcctaaca tagttaattt ttggaaggct tgccaagagg ctgcaattaa tactgtaaaa 33780 tcccgaaaga cgcatcatac acatggactt agattttata tgaaaaaagg ttttctaatg 33840 attgaactgc ctagtggaag agctttagct tatccaaaag ctttagttgg tgaaaatagt 33900 tggggtagtc aagttgttga atttatgggg ttagatctta accgtaaatg gtcaaagtta 33960 aaaacgtatg gtgggaagtt agtcgagaat attgttcaag caactgcaag ggatttactt 34020 gcgatttcta tagcaaggct tgaagcatta ggttttaaaa tagttggcca tgtccatgat 34080 gaagtaattg tagaaatacc tagaggttca aatggactta aggaaatcga aactatcatg 34140 aataagcctg ttgattgggc aaaaggattg aatttgaata gtgacgggtt tacttctccg 34200 ttttatatga aggattagga gtgtgattgc atgcaacatc aagcttatat caatgcttct 34260 gttgacatta gaattcctac agaagtcgaa agtgttaatt acaatcagat tgataaagaa 34320 aaagaaaatt tggcggacta tttatttaat aatccaggtg aactattaaa atataacgtt 34380 ataaatatta aggttttaga tttagaggtg gaatgatggc tagaagaaaa gttataagag 34440 tgcgtatcaa aggaaaacta atgacattga gagaagtttc agaaaaatat cacatatctc 34500 cagaacttct tagatataga tacaaacata aaatgcgcgg cgatgaatta ttgtgtggaa 34560 gaaaagactc aaaatctaaa gatgaagttg aatatatgca gagtcaaata aaagatgaag 34620 aaaaagagag agaaaaaatc agaaaaaaag cgattttgaa cctataccaa cgaaatgtga 34680 gagcggaata tgaagaagaa agaaagagaa gattgagacc atggctttat gatggaacgc 34740 cacaaaaaca ttcacgtgat ccgtactggt tcgatgtcac ttataaccaa atgttcaaga 34800 aatggagtga agcataatga gcgtaatcag taacagaaaa gtagatatga acgaagcgca 34860 agacaatgtt aagcaaccag cgcactacac atacggcgac attgaaatta tagattttat 34920 cgaacaggtt acggcacagt atccacctca actagcattc gcaataggta atgcaataaa 34980 atacttgtct agagcacctt taaagaatgg tcatgaggat ttagcaaagg cgaagtttta 35040 cgtccaaaga gcttttgact tgtgggagtg atgaccatga cagatagcgc atgtaaagaa 35100 tacttaaacc aatttttcgg atctaagaga tatctgtatc aggataacga acgagtggca 35160 catatccatg tagtgaatgg cacttattac tttcacgggc atatcgtacc aggctggcaa 35220 ggcgtgaaaa agacatttga tacagcggaa gagctcgaaa catatataaa gcaacatggt 35280 ttggaatacg aggaacagaa gcaactaact ttattttaag gagatagaaa tgatgaaaat 35340 caaagttgaa aaaataatga aaatagacga attaattaag tgggcgcgag aaaatccgga 35400 gctatcattt ggcagaaaat attatacaac agacaaaaat gatgaaaact ttatttactt 35460 cggtgttttt aaaaattgtt ttaaaataag cgattttata ttagttaatg ctacttttag 35520 tgtcaaagtt gaagaagaag taaccgaaga aactaagttt gataggttgt ttgaagtgta 35580 cgagattcaa gaaggagtct ataaatctgc atcatatgag aatgctagta taaacgaacg 35640 tttaaaaaat gacagaattt ttcttgctaa agcattctac atcttaaacg acgacctaac 35700 tatgacgtta atttggaaag aaggagagtt gattaaataa tggaacacgg ttcaaaagaa 35760 tattacgaaa agcaaagtga atactggttt gatgaagcaa gcaagttttt gaagcaacgt 35820 gatgagctta ttggagatat agctaagtta agagagtgca acaaagagct ggagaagaaa 35880 gcaagtgcat gggataggta ttgcaagagc gttgaaaaag atttaataaa cgaatttggc 35940 aaagatggtg aaagagttaa atttggaatg gaattaaaca ataaaatttt tatggaggaa 36000 gacgcaaatg aataaccgcg aacaaatcga acaatcagtt attagtgcta gcgcgtataa 36060 cggcaatgac acagagggat tattaaaaga gattgaggac gtgtataaga aagcgcaagc 36120 gtttgatgaa atacttgagg gtttacctaa tgctatgcaa gatgcaatca aagaagatat 36180 tggtcttgat gaagcagtag gaattatgac gggtcaagtt gtctataaat atgaggagga 36240 gcaggaaaat gactaacata ttacaagtga aactattatc aaaagacgct agaatgccag 36300 aacgaaatca taagacggat gcaggttatg acatattttc agctaaaact gtcgtacttg 36360 agccacaaga aaaggcagtg atcaaaacag atgtagctgt aagcattcca gagggctatg 36420 tcggtttatt aactagccgt agtggtgtaa gtagtaaaac gcatttagtg attgaaacag 36480 gcaagataga cgcgggatat catggtaatt tagggattaa tatcaagaat gataatgaaa 36540 cgttagagag tgaggatatg agtaactttg gtcggagtcc ttctggtata gatggaaaat 36600 acaccctact acctgtaaca gataaatttt tatgtatgaa tggtagttat gtcataaata 36660 aaggcgacaa actagctcaa ttggttatcg tgcctatatg gacacctgaa ctaaagcaag 36720 tggaggaatt cgagagtgtt tcagaacgtg gagcaaaagg cttcggaagt agcggagtgt 36780 aaagacatat tagatcgagt caaggaggtt ttggggaagt gagtgacatg ttagaaatat 36840 ttttcatagg gtttggtgtt tatctatttt gtcgcatagg tattattttt ctcaagagta 36900 aaaagactat acacacaaac ctatatgaaa tgttgttgat tgctactatc tttgtgacat 36960 ctacatttgc tgataaacat caaaagacgc atatcttaat agcattttta gtaatgtttt 37020 ttatgagtaa gctcaaacaa gttcaaggga gctatgagga atgacacaat acctagtcac 37080 aacatttaaa gattcaacag gacgtaagca tacacacata actaaagcta agagcaatca 37140 aaggtttaca gttgttgatg cggagagtaa agaagaagcg aaagagaagt acgaggcaca 37200 agttaaaaga aatgcagtta ttaaattagg gcagttgttt gaaaatataa gggagtgtgg 37260 gaaatgacta aacaaatact aagattatta ttcttactag cgatgtatga gctaggcaag 37320 tatgtaactg agcaagtata tattatgatg acggctaatg atgatgcaga ggcgccgagt 37380 gactttgaaa aaatcagagc tgaagtttca tggtaatagc tattatcatt tttgaattaa 37440 ttatattaat gtgtttagca atagcactgg aggtgttgta aatatgtgga ttgtcatttc 37500 aattgtttta tctatatttt tattgatctt gttaagtagc atttctcata agatgaaaac 37560 catagaagca ttggagtata tgaatgctta tcttttcaag cagttagtaa aaaataatgg 37620 tgttgaaggt atagaagatt atgaaaatga agttgaacga attagaaaaa gatttaaaag 37680 ctaaagagag gcgttggctt ctctgttcta tttaaaataa tgaaaggagc cgaacatgtt 37740 agacaaagtc actcaaatag aaacaattaa atatgatcgt gatgtttcat attcttatgc 37800 tgctagtcgt ttatctacac attggactaa tcacaatatg gcttggtctg actttatgca 37860 gaagctagca caaacagtta gaactaaaga agatttaact gagtacaata aaatgtctaa 37920 gtctgaacaa gccgatataa aagatgttgg cggatttgtc ggtggttatt taaaagaagg 37980 caaacgacgt gctggtcaag tcatgaatcg ttcaatgtta acacttgata tcgattatgc 38040 tgctcaagat atgactgaca tattatctat gttttatgat tttgcatatt gtttatattc 38100 aacacataag catagagaga taagtccaag actgcgttta gtgattcctt taaaacgaaa 38160 tgtaaatgca gatgagtatg aagctattgg gcgtaaagtc gcagatatcg ttggcatgga 38220 ttacttcgat gatacaactt atcaaccaca taggttaatg tattggcctt caactagtaa 38280 cgatgcggaa tttttcttta cctatgaaga tttacctttg ttagacccag ataaaatatt 38340 aaatgaatat gttgattgga ctgacacatt agaatggcca acgtcttcaa gggaagagag 38400 taagactaaa agattagcag ataagcaagg cgacccagaa gaaaagccgg gaattgttgg 38460 tgcattttgt agagcctata cgatagaaga agctatagaa acttttattc ctgatttata 38520 cgaaaaacat tctactaacc gttataccta tcatgaaggt tcaactgcag gtggattggt 38580 gttatacgaa aataacaagt ttgcctattc tcatcataat acggatcccg taagcggtat 38640 gcttgtgaac agttttgatt tagtacgcat acacttatat ggtgctcaag atgaagacgc 38700 taaaacagat actccggtta atcgactacc tagttataaa gcaatgcagc aaagagcgca 38760 aaatgatgaa gttgttaaaa agcaattaat taacgacaaa atgtctgatg caatgcagga 38820 tttcgatgaa atagtaaata gcgatgatgc atggtctgag acgttagaaa ttacttcgaa 38880 aggtactttc aaagctagta tcccaaatat agaaattata ttgcgtaatg atccaaattt 38940 aaaaggaaaa atagcattta atgaatttac aaaacaaatt gaatgcttag ggaaaatgcc 39000 atggaataat aattttaaaa tacgtcaatg gcaagacggt gatgatagca gtttaagaag 39060 ttatatcgaa aagatttatg acatacacca ttcaggcaaa acaaaagatg ccattataag 39120 cgtagcaatg caaaatgcct atcatccagt aagagattat ctaaataaaa tatcgtggga 39180 tggacataaa cgtcttgaaa agttatttat caaatactta ggtgttgaag acactgaagt 39240 gaatagaaca actaccaaaa aggcattgac tgctggaatc gctcgagtaa tggagccagg 39300 atgtaaattt gactatatgc ttacacttta tggtcctcaa ggtgtaggta aatctgcttt 39360 gctaaaaaaa ataggtggtg catggttttc tgacagttta gtttctgtta ctggtaagga 39420 agcatatgag gcattacaag gcgtttggtt aatggaaatg gcagaacttg cagctacaag 39480 aaaagctgaa gttgaagcta ttaagcattt catatctaaa caagttgacc ggtttcgtgt 39540 tgcttatgga cattatattg aagattttcc aaggcaatgt attttcattg gtacaactaa 39600 taaagttgat ttcttaagag atgaaactgg tggaagacgt ttttggccaa tgactgtaaa 39660 tccagagaga gttgaagtga actggtctaa actaaccaaa gaagagatcg accaaatctg 39720 ggcagaagct aaatactatt atgaacaagg agaagagttg ttccttaacc ctgaactaga 39780 agaagaaatg cgttcaatcc aaagtaaaca tactgaggaa tctccatata caggtattat 39840 tgatgaatat cttaacacgc caatcccaag caattgggaa gacttaacta tctttgaaag 39900 aagacgattt tatcaaggtg atgttgatat gttaccaaca ggaaatgtag attacattga 39960 aagagacaag gtctgtgcgc ttgaagtgtt tgttgaatgt tttggtaaag ataagggaga 40020 tagtagagga tctatggaaa ttagaaagat ttctaacgtc ttaagacaat tagacaattg 40080 gtctgtatat gaaggcaata aaagtgggaa aattcgattt ggaaaagatt atggtgtaca 40140 gatagcgtat gtaagagatg aaagtttaga ggatttaata taagaaatat tgaataaata 40200 tacattttta gatgttgtat caaatgttgc atcatttttt gagtgatgca acacggtggt 40260 gtaaaaagta atcgtaggtg ttgtatcatt tttggtgatg caacattgat gcaacaaatg 40320 atacaacacc tctttccctt ctcgctgtaa ggttcaaccc tgtttgtttc caatgttgca 40380 tcaaattcac tataaagttt aaaaagtagt gttagggagt aaaggggtat aggggtaacc 40440 ctctaacagc tatttttaaa agtttggcaa gaattgatgc aacatcggaa cacaaatata 40500 aattttgtat acaaggtgaa taaatgaaag aatcgacatt agaaaaatat ttagtgaaag 40560 agataacaaa gttaaatgga ttatgtttaa aatgggtcgc acctggaaca agaggtgtac 40620 cagatagaat tattattatg ccagaaggaa aaacatattt tgtagaaatg aagcaagaaa 40680 agggaaagtt acatccttta caaaaatatg tgcatcggca atttgaaaac agagatcata 40740 cagtgtatgt gttatggaat aaagaacaag taaatacttt tataagaatg gtaggtggaa 40800 catttggcga ttgatttcaa accacatagc tatcaaaagt atgcaataga taaagtgatt 40860 gataatgaga aatacggttt gtttttagat atggggctag ggaaaacagt atcaacactt 40920 acagcattta gtgaattgca gttgttagac actaaaaaaa tgttagtcat agcacctaaa 40980 caagttgcta aagatacatg ggttgatgaa gttgataagt ggaaccattt aaatcatctg 41040 aaagtgtctt tagtcttagg aacacctaaa gaaagaaatg atgcattaaa cacagaggct 41100 gatatctatg taaccaataa agaaaatact aaatggttat gtgatcaata taaaaaagaa 41160 tggccatttg acatggttgt aattgatgaa ctgtctacat ttaaaagtcc taagagtcaa 41220 aggtttaaat ctattaaaaa gaaattacca ctcattaata gatttatagg attaacagga 41280 acacctagtc caaatagttt acaggattta tgggctcaag tttatttgat agacagaggc 41340 gaaagacttg agtcttcatt cagtcgttat cgagaaaggt actttaaacc aacacatcaa 41400 gttagcgaac atgtttttaa ctgggagcta agagacggat ctgaagaaaa gatatatgaa 41460 cgaatagaag atatatgttt aagcatgaaa gcgaaagatt atctggatat gcctgacaga 41520 gttgatacta aacaaacagt agtcttatct gaaaaagaaa gaaaagtata tgaagaatta 41580 gaaaaaaact atattttaga atcggaagaa gaaggaacag ttgtagctca gaatggggca 41640 tcattaagtc aaaaactact tcaactatct aacggtgcag tttatacaga tgatgaagat 41700 gtaagactta tacatgataa gaagttagat aagttagagg aaattataga ggagtctcaa 41760 ggccaaccaa tattattgtt ttataacttc aaacatgata aagaaagaat acttcaaagg 41820 tttaaggaag caaccacatt agaggattca aactataaag aacgttggaa tagtggagac 41880 attaagctgc ttatagcaca tccagcaagt gcagggcatg gattaaactt acaacaaggt 41940 gggcacatta ttgtttggtt tggacttaca tggtcattgg aattatacca acaagcaaat 42000 gcaagattat atagacaagg acaaaatcat acgactatta ttcatcacat catgaccgat 42060 aacacaatag atcaaagagt atataaagct ttacaaaata aagaactaac gcaagaagaa 42120 ttgatgaaag ctattaaagc aagaatagct aagcataagt aatggaggta taagatggga 42180 aaggcgtcat atgatattaa gccaggaaca tttaaatata ttgaatcaga aatatataat 42240 ttaaatgaga acaagaaaga gataaataga ttgagaatgg agatacttaa cccaacgaaa 42300 gaactagaca ccaacattgt gtatggaccg ttacaaaaag gagagccagt tagaacaact 42360 gagttaatgg cgacaaggtt attgactaat aagatgttac gtaacttaga agagatggtt 42420 gaagcagttg aaagtgagta cttaaagtta cctgaagatc ataagaaagt aataaggtta 42480 aagtattgga ataaagataa gaagctaaag atagaacaaa taggggatgc ttgtcacatg 42540 catcgcaata cagttactac aatacgaaag aactttgtta aagcgatagc gtatcatgca 42600 ggtatcaaat aacattgtgc aaagattgtg caaaaggcct acaaatctgt agtaatatga 42660 tagtatcgga aagatgtata aagttatctg aaagttatac gacataaata catgaggcac 42720 atcgctaagc ggtgtgtctt ttgttatgca atcaaagagg tgtaagagat gaccaagcat 42780 aataacattt ataagcatgg tcgtaagtca tatcaatacg attggttcta tcattcaaaa 42840 gcatggaaga agttaagaga gatagcatta gatagagata attatctttg tcaaatgtgt 42900 ttacgcgaag atattataac agatgcaaag attgtgcatc acattattta tgttgatgaa 42960 gattttaaca aagctttaga cttagataat ctaatgtcag tttgttatag ctgtcataac 43020 aaaattcatg caaatgataa tgacaaaagt aatcttaaga aaattagagt tctaaaaatt 43080 taaataaaaa aatta 43095 18 41708 DNA Staphylococcus bacteriophage 18 gatcaaaata cttggggaac ggttagggag taaacttcgc gataatttta aaaattcatg 60 tataaccccc ctcttataac cattttaagg caggtgatga aatggagatt atagtcgatg 120 aaaatttagt gcttaaagaa aaagaaaggc tacaagtatt atataaagac atacctagca 180 ataaattaaa agtagttgat ggtttaatta ttcaagcagc aaggctacgt gtaatgcttg 240 attacatgtg ggaagacata aaagaaaaag gtgattatga tttatttact caatctgaaa 300 aggcgccacc atatgaaagg gaaagaccag tagccaaact atttaatgct agagatgctg 360 catatcaaaa aataatcaaa caattatcgg atttattgcc cgaagagaaa gaagacacag 420 aaacgccatc tgatgattac ctatgattag taataaatac gttgatgaat atataaattt 480 gtggaaacaa ggaaagataa ttttaaataa agaaagaatt gatctcttta attatctaca 540 aaaacatata tattcacgag atgatgtata ttttgatgaa cagaaaatcg aggattgtat 600 caaatttatt gaaaaatggt attttccaac attaccattt caaaggttta tcatagctaa 660 tatatttctt atagataaaa atacagatga agctttcttt acagaatttg ctattttcat 720 gggacgtgga ggcgggaaaa acggtctaat aagtgctatt agtgattttc tttctacgcc 780 cttacacgga gttaaagaat atcacatctc cattgttgct aatagtgaag atcaagcaaa 840 aacatcgttt gatgaaatca gaaccgtttt aatggataac aaacgaaata agacgggtaa 900 aacgccaaaa gctccttatg aagttagtaa agcaaaaata ataaaccgtg caactaaatc 960 ggttattcga tataacacat caaacacaaa aaccaaagac ggtggacgtg aggggtgtgt 1020 tatttttgat gaaattcatt atttctttgg tcctgaaatg gtaaacgtca aacgtggtgg 1080 attaggtaaa aagaaaaata gaagaacgtt ttatataagt actgatggtt ttgttagaga 1140 gggttatatc gatgcaatga agcacaaaat tgcaagtgta ttaagtggca aggttaaaaa 1200 tagtagattg tttgcttttt attgtaagtt agacgatcca aaagaagttg atgacagaca 1260 gacgtgggaa aaggcgaacc caatgttaca taaaccgtta tcagaatacg ctaaaacact 1320 gctaagcacg attgaagaag aatataacga tttaccattc aaccgttcaa ataagcccga 1380 attcatgact aagcgaatga atttgcctga agttgacctt gaaaaagtaa tagcaccatg 1440 gaaagaaata ctagcgacta atagagagat accaaattta gataatcaaa tgtgtattgg 1500 tggtttagac tttgcaaaca ttcgagattt tgcaagtgta gggctattat tccgaaaaaa 1560 cgatgattac atttggttag gacattcgtt tgtaagacaa gggtttttgg atgatgtcaa 1620 attagaacct cctattaaag aatgggaaaa aatgggatta ttgaccattg tcgatgatga 1680 tgtcattgaa attgaatata tagttgattg gtttttaaag gctagagaaa aatatgggct 1740 tgaaaaagtc atagctgata attatagaac tgatattgta agacgtgcgt ttgaggatgc 1800 tggcataaaa cttgaagtac ttagaaatcc aaaagcaata catggattac ttgcaccacg 1860 tatcgataca atgtttgcga aacataacgt aatatatgga gacaatcctt tgatgcgttg 1920 gtttactaat aatgttgctg taaaaatcaa gccggatgga aataaagagt atatcaaaaa 1980 agatgaagtc agacgtaaaa cggatggatt catggctttt gttcacgcat tatatagagc 2040 agacgatata gtagacaaag acatgtctaa agcgcttgat gcattaatga gtatagattt 2100 ctaatagagg aggtgagaca tgagtattct agaaaagata tttaaaacta ggaaagatat 2160 aacatatatg cttgatttag atatgataga agatctatca caacaagcgt atgtgaaacg 2220 tttagcgatt gatagttgta ttgaatttgt tgcgcgagct gtcgctcaaa gtcattttaa 2280 agtattggaa ggtaatagaa ttcaaaagaa tgatgtttac tacaagttaa atataaaacc 2340 aaatactgac ttatcaagcg atagtttttg gcaacaagtt atatataaac taatttatga 2400 taacgaggtt ttaatcgtag taagtgacag caaagaatta cttatcgcag atagctttta 2460 cagagaagag tacgctttgt atgatgatat attcaaagat gtaacggtta aagattatac 2520 ttatcaacgt actttcacaa tgcaagaggt catatattta aagtacaaca acaataaagt 2580 gacacacttt gtagaaagtc tattcgaaga ttacgggaaa atattcggaa gaatgatagg 2640 tgcacaatta aaaaactatc aaataagagg gattttgaaa tctgcctcta gcgcatatga 2700 cgaaaagaat atagaaaaat tacaagcgtt cacaaataaa ttattcaata cttttaataa 2760 aaatcaacta gcaatcgcgc ctttgataga aggttttgat tatgaggaat tatctaatgg 2820 tggtaagaat agtaacatgc ctttttctga attgagtgag ctaatgagag atgcaataaa 2880 aaatgttgcg ttgatgattg gtatacctcc aggtttgatt tacggagaaa cagctgattt 2940 ggaaaaaaac acgcttgtat ttgagaagtt ctgtttaaca cctttattaa aaaagattca 3000 gaacgaatta aacgcgaaac tcataacaca aagcatgtat ttgaaagata caagaataga 3060 aattgtcggt gtgaataaaa aagacccact tcaatatgct gaagcaattg acaaacttgt 3120 aagttctggt tcatttacaa ggaatgaggt gcggattatg ttaggtgaag aaccatcaga 3180 caatcctgaa ttagacgaat acctgattac taaaaactac gaaaaagcta acagtggtga 3240 aaatgatgaa aaagaaaaag atgaaaacac tttgaaaggt ggtgatgaag atgaaagcgg 3300 agattaaagg cgtcatcgtt tccaacgaag ataaatgggt ttacgaaatg cttggtatgg 3360 attcgacttg tcctaaagat gttttaacac aactagaatt tagtgatgaa gatgttgata 3420 ttataattaa ctcaaatggt ggtaacctag tagctggtag tgaaatatat acacatttaa 3480 gagctcataa aggcaaagtg aatgttcgta tcacagcaat agcagcaagt gcggcatcgc 3540 ttatcgcaat ggctggtgac cacatcgaaa tgagtccggt tgctagaatg atgattcaca 3600 atccttcaag tattgcgcaa ggagaagtga aagatctaaa tcatgctgca gaaacattag 3660 aacatgttgg tcaaataatg gctgaggcat atgcggttag agctggtaaa aacaaacaag 3720 aacttataga aatgatggct aaggaaacgt ggctaaatgc tgatgaagcc attgaacaag 3780 gttttgcgga tagtaaaatg tttgaaaacg acaatatgca aattgtagca agcgatacac 3840 aagtgttatc gaaagatgta ttaaatcgtg taacagcttt ggtaagtaaa acgccagagg 3900 ttaacattga tattgacgca atagcaaata aagtaattga aaaaataaat atgaaagaaa 3960 aggaatcaga aatcgatgtt gcagatagta aattatcagc aaatggattt tcaagattcc 4020 ttttttaata caaaaatagg aggtcataaa atgactataa atttatcgga aacattcgca 4080 aatgcgaaaa acgaatttat taatgcagta aacaacggtg aaccgcaaga aagacaaaat 4140 gaattgtacg gtgacatgat taaccaacta tttgaagaaa ctaaattaca agcaaaagca 4200 gaagctgaaa gagtttctag tttacctaaa tcagcacaaa ctttgagtgc aaaccaaaga 4260 aatttcttta tggatatcaa taagagtgtt ggatataaag aagaaaaact tttaccagaa 4320 gaaacaattg atagaatctt cgaagattta acaacgaatc atccattatt agctgactta 4380 ggtattaaaa atgctggttt gcgtttgaag ttcttaaaat ccgaaacttc tggcgtggct 4440 gtttggggta aaatctatgg tgaaattaaa ggtcaattag atgctgcgtt cagtgaagaa 4500 acagcaattc aaaataaatt gacagcgttt gttgttttac caaaagattt aaatgatttt 4560 ggtcctgcgt ggattgaaag atttgttcgt gttcaaatcg aagaagcatt tgcagtggcg 4620 cttgaaactg cgttcttaaa aggtactggt aaagaccaac cgattggctt aaaccgtcaa 4680 gtacaaaaag gtgtatcggt aactgatggt gcttatccag agaaagaaga acaaggtacg 4740 cttacatttg ctaatccgcg cgctacggtt aatgaattga cgcaagtgtt taaataccac 4800 tcaactaacg agaaaggtaa atcagtagcg gttaaaggta atgtaacaat ggttgttaat 4860 ccgtccgatg cttttgaggt tcaagcacag tatacacatt taaatgcaaa tggcgtatat 4920 gttactgctt taccatttaa tttgaatgtt attgagtcta cagttcaaga agcaggtaag 4980 gttttaacgt acgttaaagg tctatatgat ggttatttag ctggtggtat taatgttcag 5040 aaatttaaag aaacacttgc gttagatgat atggatttat acactgcaaa acaatttgct 5100 tacggcaaag cgaaagataa taaagttgct gctgtttgga aattagattt aaaaggacat 5160 aaaccagctt tagaagatac cgaagaaaca ctataaaatt ttatgaggtg ataaaatggt 5220 gaaatttaaa gttgttagag aatttaaaga catagagcac aatcaacaca agtacaaagt 5280 aggggagttg tatccagctg aagggtataa caatcctcgt gttgaattgt tgacaaatca 5340 aatcaaaaat aagtacgaca aagtttatat cgtaccttta gataagctga caaaacaaga 5400 attattagaa ctatgcgaat cattacaaaa aaaagcgtct agttcaatgg ttaaaagtga 5460 aatcatcgac ttattgaatg gtgaagacaa tgacgattga tgatttgctt gtcaaattta 5520 aatcacttga aaagattgac cataattcag aggatgagta cttaaagcag ttgttaaaaa 5580 tgtcgtacga gcgtataaaa aatcagtgcg gagtttttga attagagaat ttaataggtc 5640 aagaattgat acttatacgc gctagatatg cttatcaaga tttattagaa cacttcaacg 5700 acaattacag acctgaaata atagattttt cgttatctct aatggaggta tcagaagatg 5760 aagaaagtgt ttaagaaacc tagaattaca actaaacgtt taaatacgcg tgttcatttt 5820 tataagtata ctgaaaataa tggtccagaa gctggagaaa aagaagaaaa attattatat 5880 agctgttggg cgagtattga tggtgtctgg ttacgtgaat tagaacaagc tatctcaaac 5940 ggaacgcaaa atgacattaa attgtatatt cgtgatccgc aaggtgatta tttacccagt 6000 gaagaacatt atcttgaaat tgaatcaaga tatttcaaaa atcgtttgaa tataaagcaa 6060 gtatcaccag atttggataa taaagacttt attatgattc gcggaggata tagttcatga 6120 gtgtgaaagt gacaggtgat aaagcattag aaagagaatt agaaaaacat tttggcataa 6180 aagagatggt aaaagttcaa gataaggcgt taatagctgg tgctaaggta attgttgaag 6240 aaataaaaaa acaactcaaa ccttcagaag actcaggagc actgattagt gagattggtc 6300 gtactgaacc tgaatggata aaggggaaac gtactgttac aattaggtgg cgtgggcctt 6360 ttgaacgatt tagaatagta catttaattg aaaatggtca tgttgagaaa aagtcaggaa 6420 aatttgtaaa acctaaagct atgggtggga ttaatagagc aataagacaa gggcaaaata 6480 agtattttga gacgctaaaa agggagttga aaaaattgtg attgatattt tgtacaaagt 6540 tcatgaagtg attagtcaag acagaattat tagagagcac gtaaatatca ataatattaa 6600 gttcaataaa taccctaatg taaaagatac tgatgtacct tttattgtta ttgacgatat 6660 cgacgaccca atacctacaa cttatactga cggagatgag tgtgcatata gttatattgt 6720 ccaaatagat gtttttgtta agtacaatga tgaatataat gcgagaatca taagaaataa 6780 gatatctaat cgcattcaaa agttattatg gtctgaacta aaaatgggaa atgtttcaaa 6840 tggaaaaccg gaatatatag aagaatttaa aacatataga agctctcgcg tttacgaggg 6900 cattttttat aaggaggaaa attaaatggc agtaaaacat gcaagtgcgc caaaggcgta 6960 tattaacatt actggtttag gtttcgctaa attaacgaaa gaaggcgcgg aattaaaata 7020 tagtgatatt acaaaaacaa gaggattaca aaaaattggt gttgaaactg gtggagaact 7080 aaaaacagct tatgctgatg gcggtccaat tgaatcaggg aatacagacg gagaaggtaa 7140 aatctcatta caaatgcatg cgttccctaa agagattcgc aaaattgttt ttaatgaaga 7200 ttatgatgaa gatggcgttt acgaagagaa acaaggtaaa caaaacaatt acgtagctgt 7260 atggttcaga caagagcgta aagacggtac atttagaaca gttttattac ctaaagttat 7320 gtttacaaat cctaaaatcg atggagaaac ggctgagaaa gattgggatt tctcaagtga 7380 agaggttgaa ggtgaggcac ttttcccttt agttgataat aaaaagtcag tacgtaagta 7440 tatctttgat tcagctaaca tgacaaatca tgatggagac ggtgaaaaag gcgaagaggc 7500 tttcttaaag aaaattttag gcgaagaata tactggaaac gtgacagagg gtaacgaaga 7560 aactttgtaa caaaaccggc ttcatcggaa actgcggtaa agtcggttaa tataccagat 7620 agcattaaaa cacttaaagt tggcgacaca tacgatttaa atgttgtagt agagccatct 7680 aatcaaagta agttattgaa atacacaaca gatcaaacga atattgtatc aatcaatagt 7740 gatggtcaag ttactgcgga agcacaaggc attgctacgg ttaaagcaac agttggtaat 7800 atgagtgaca ctataacaat aaatgtagaa gcataagagg gggcaacccc tctattttat 7860 ttgaaaataa ggagagtatt ataaaatggc aaaattaaaa cgtaacatta ttcaattagt 7920 agaagatcca aaagcaaatg aaattaaatt acaaacgtac ttaacaccac acttcatttc 7980 atttgaaatt gtatacgaag caatggattt aatcgatgat attgaggacg aaaatagcac 8040 gatgaagcca agagaaatcg ctgacagatt gatggatatg gttgtaaaaa tttacgataa 8100 ccaattcaca gttaaagacc taaaagaacg tatgcatgca cctgatggaa tgaatgcact 8160 tcgtgaacaa gtgattttca ttactcaagg tcaacaaact gaggaaacta gaaattttat 8220 ccagaacatg aaataaagcc tgaagattta acatataaag caatgttgaa aaatatggat 8280 actctcatga tggacttaat tgaaaatggt aaagacgcta acgaagtttt aaaaatgcca 8340 tttcattatg tgctttccat atatcaaaat aaaaataatg acatttctga agaaaaagca 8400 gaggctttaa ttgatgcatt ttaaccttaa ccgtttggtt agggttattt ttttgaactt 8460 ttttagaaag gaggtaaaaa atgggagaaa gaataaaagg tttatctata ggtttggatt 8520 tagatgcagc aaatttaaat agatcatttg cagaaatcaa acgaaacttt aaaactttaa 8580 attctgactt aaaattaaca ggcaacaact tcaaatatac cgaaaaatca actgatagtt 8640 acaaacaaag gattaaagaa cttgatggaa ctatcacagg ttataagaaa aacgttgatg 8700 atttagccaa gcaatatgac aaggtatctc aagaacaggg cgaaaacagt gcagaagctc 8760 aaaagttacg acaagaatat aacaaacaag caaatgagct gaattattta gaaagagaat 8820 tacaaaaaac atcagccgaa tttgaagagt tcaaaaaagc tcaagttgaa gctcaaagaa 8880 tggcagaaag tggctgggga aaaaccagta aagtttttga aagtatggga cctaaattaa 8940 caaaaatggg tgatggttta aaatccattg gtaaaggttt gatgattggt gtaactgcac 9000 ctgttttagg tattgcagca gcatcaggaa aagcttttgc agaagttgat aaaggtttag 9060 atactgttac tcaagcaaca ggcgcaacag gcagtgaatt aaaaaaattg cagaactcat 9120 ttaaagatgt ttatggcaat tttccagcag atgctgaaac tgttggtgga gttttaggag 9180 aagttaatac aaggttaggt tttacaggta aagaacttga aaatgccaca gagtcattct 9240 tgaaattcag tcatataaca ggttctgacg gtgtgcaagc cgtacagtta attacccgtg 9300 caatgggcga tgcaggtatc gaagcaagtg aatatcaaag tgttttggat atggtagcaa 9360 aagcggcgca agctagtggg ataagtgttg atacattagc tgatagtatt actaaatacg 9420 gcgctccaat gagagctatg ggctttgaga tgaaagaatc aattgcttta ttctctcaat 9480 gggaaaagtc aggcgttaat actgaaatag cattcagtgg tttgaaaaaa gctatatcaa 9540 attggggtaa agctggtaaa aacccaagag aagaatttaa gaagacatta gcagaaattg 9600 aaaagacgcc ggatatagct agcgcaacaa gtttagcgat tgaagcattt ggtgcaaagg 9660 caggtcctga tttagcagac gctattaaag gtggtcgctt tagttatcaa gaatttttaa 9720 aaactattga agattcccaa ggcacagtaa accaaacatt taaagattct gaaagtggct 9780 ccgaaagatt taaagtagca atgaataaat taaaattagt aggtgctgat gtatgggctt 9840 ctattgaaag tgcgtttgct cccgtaatgg aagaattaat caaaaagcta tctatagcgg 9900 ttgattggtt ttccaattta agtgatggtt ctaaaagatc aattgttatt ttcagtggta 9960 ttgctgctgc aattggtcct gtagtttttg ggttaggtgc atttataagt acaattggca 10020 atgcagtaac tgtattagct ccattgttag ctagtattgc aaaggctggt ggattgatta 10080 gttttttatc gactaaagta cctatattag gaactgtctt cacagcttta actggtccaa 10140 ttggcattgt attaggtgta ttggctggtt tagcagtcgc atttacaatt gcttataaga 10200 aatctgaaac atttagaaat tttgttaatg gtgcaattga aagtgttaaa caaacattta 10260 gtaattttat tcaatttatt caacctttcg ttgattctgt taaaaacatc tttaaacaag 10320 cgatatcagc aatagttgat ttcgcaaaag atatttggag tcaaatcaat ggattcttta 10380 atgaaaacgg aatttccatt gttcaagcac ttcaaaatat atgcaacttt attaaagcga 10440 tatttgaatt tattttaaat tttgtaatta aaccaattat gttcgcgatt tggcaagtga 10500 tgcaatttat ttggccggcg gttaaagcct tgattgtcag tacttgggag aacataaaag 10560 gtgtaataca aggtgcttta aatatcatac ttggcttgat taagttcttc tcaagtttat 10620 tcgttggtga ttggcgagga gtttgggacg ccgttgtgat gattcttaaa ggagcagttc 10680 aattaatttg gaatttagtt caattatggt ttgtaggtaa aatacttggt gttgttaggt 10740 actttggcgg gttgctaaaa ggattgatag caggaatttg ggacgtaata agaagtatat 10800 tcagtaaatc tttatcagca atttggaatg caacaaaaag tatttttgga tttttattta 10860 atagcgtaaa atcaattttc acaaatatga aaaattggtt atctaatact tggagcagta 10920 tccgtacgaa tacaatagga aaagcgcagt cattatttag tggcgtcaaa tcaaaattta 10980 ctaatttatg gaatgcgacg aaagaaattt ttagtaattt aagaaattgg atgtcaaata 11040 tttggaattc cattaaagat aatacggtag gaattgcaag ccgtttatgg agtaaggtac 11100 gtggaatttt cacaaatatg cgcgatggct tgagttccat tatagataag attaaaagtc 11160 atatcggcgg tatggtaagc gctattaaaa aaggacttaa taaattaatc gacggtttaa 11220 actgggtcgg tggtaagttg ggaatggata aaatacctaa gttacacact ggtacagagc 11280 acacacatac tactacaaga ttagttaaga acggtaagat tgcacgtgac acattcgcta 11340 cagttgggga taagggacgc ggaaatggtc caaatggttt tagaaatgaa atgattgaat 11400 tccctaacgg taaacgtgta atcacaccta atacagatac taccgcttat ttacctaaag 11460 gctcaaaagt atacaacggt gcacaaactt attcaatgtt aaacggaacg cttccaagat 11520 ttagtttagg tactatgtgg aaagatatta aatctggtgc atcatcggca tttaactgga 11580 caaaagataa aataggtaaa ggtaccaaat ggcttggcga taaagttggc gatgttttag 11640 attttatgga aaatccaggc aaacttttaa attatatact tgaagctttt ggaattgatt 11700 tcaattcttt aactaaaggt atgggaattg caggcgacat aacaaaagct gcatggtcta 11760 agattaagaa aagtgctact gattggataa aagaaaattt agaagctatg ggcggtggcg 11820 atttagtcgg cggaatatta gaccctgaca aaattaatta tcattatgga cgtaccgcag 11880 cttataccgc tgcaactgga agaccatttc atgaaggtgt cgattttcca tttgtatatc 11940 aagaagttag aacgccgatg ggtggcagac ttacaagaat gccatttatg tctggtggtt 12000 atggtaatta tgtaaaaatt actagtggcg ttatcgatat gctatttgcg catttgaaaa 12060 actttagcaa atcaccacct agtggcacga tggtaaagcc cggtgatgtt gttggtttaa 12120 ctggtaatac cggatttagt acaggaccac atttacattt tgaaatgagg agaaatggac 12180 gacattttga ccctgaacca tatttaagga atgctaagaa aaaaggaaga ttatcaatag 12240 gtggtggcgg tgctacttct ggaagtggcg caacttatgc cagtcgagta atccgacaag 12300 cgcaaagtat tttaggtggt cgttataaag gtaaatggat tcatgaccaa atgatgcgcg 12360 ttgcaaaacg tgaaagtaac taccagtcaa atgcagtgaa taactgggat ataaatgctc 12420 aaagaggaga cccatcaaga ggattattcc aaatcatcgg ctcaactttt agagcaaacg 12480 ctaaacgtgg atatactaac tttaataatc cagtacatca aggtatctca gcaatgcagt 12540 acattgttag acgatatggt tggggtggtt ttaaacgtgc tggtgattac gcatatgcta 12600 caggtggaaa agtttttgat ggttggtata acttaggtga agacggtcat ccagaatgga 12660 ttattccaac agatccagct cgtagaaatg atgcaatgaa gattttgcat tatgcagcag 12720 cagaagtaag agggaaaaaa gcgagtaaaa ataagcgtcc tagccaatta tcagacttaa 12780 acgggtttga tgatcctagc ttattattga aaatgattga acaacagcaa caacaaatag 12840 ctttattact gaaaatagca caatctaacg atgtgattgc agataaagat tatcagccga 12900 ttattgacga atacgctttt gataaaaagg tgaacgcgtc tatagaaaag cgagaaaggc 12960 aagaatcaac aaaagtaaag tttagaaaag gaggaattgc tattcaatga tagacactat 13020 taaagtgaac aacaaaacaa ttccttggtt gtatgtcgaa agagggtttg aaataccctc 13080 ttttaattat gttttaaaaa cagaaaatgt agatggacgt tcggggtcta tatataaagg 13140 gcgtaggctt gaatcttata gttttgatat acctttggtg gtacgtaatg actatttatc 13200 tcacaacggc attaaaacac atgatgacgt cttgaatgaa ttagtaaagt tttttaacta 13260 cgaggaacaa gttaaattac aattcaaatc taaagattgg tactggaacg cttatttcga 13320 aggaccaata aagctgcaca aagaatttac aatacctgtt aagttcacta tcaaagtagt 13380 actaacagac ccttacaaat attcagtaac aggaaataaa aatactgcga tttcagacca 13440 agtttcagtt gtaaatagtg ggactgctga cactccttta attgttgaag cccgagcaat 13500 taaaccatct agttacttta tgattactaa aaatgatgaa gattatttta tggttggtga 13560 tgatgaggta accaaagaag ttaaggatta catgcctcct gtttatcata gtgagtttcg 13620 tgatttcaaa ggttggacta agatgattac tgaagatatt ccaagtaatg acttaggtgg 13680 taaggtcggc ggtgactttg tgatatccaa tcttggcgaa ggatataaag caactaattt 13740 tcctgatgca aaaggttggg ttggtgctgg cacgaaacga gggctcccta aagcgatgac 13800 agattttcaa attacctata aatgtattgt tgaacaaaaa ggtaaaggtg ccggaagaac 13860 agcacaacat atttatgata gtgatggtaa gttacttgct tctattggtt atgaaaataa 13920 atatcatgat agaaaaatag gacatattgt tgttacgttg tataaccaaa aaggagaccc 13980 caaaaagata tacgactatc agaataaacc gataatgtat aacttggaca gaatcgttgt 14040 ttatatgcgg ctcagaagag taggtaataa attttctatt aaaacttgga aatttgatca 14100 cattaaagac ccagatagac gtaaacctat tgatatggat gagaaagagt ggatagatgg 14160 cggtaagttt tatcagcgtc cagcttctat catagctgtc tatagtgcga agtataacgg 14220 ttataagtgg atggagatga atgggttagg ttcattcaat acggagattc taccgaaacc 14280 gaaaggcgca agggatgtca ttatacaaaa aggtgattta gtaaaaatag atatgcaagc 14340 aaaaagtgtt gtcatcaatg aggaaccaat gttgagcgag aaatcgtttg gaagtaatta 14400 tttcaatgtt gattctgggt acagtgaatt aatcatacaa cctgaaaacg tctttgatac 14460 gacggttaaa tggcaagata gatatttata gaaaggagat gagagtgtga tacatgtttt 14520 agattttaac gacaagatta tagatttcct ttctactgat gacccttcct tagttagagc 14580 gattcataaa cgtaatgtta atgacaattc agaaatgctt gaactgctca tatcatcaga 14640 aagagctgaa aagttccgtg aacgacatcg tgttattata agggattcaa acaaacaatg 14700 gcgtgaattt attattaact gggttcaaga tacgatggac ggctacacag agatagaatg 14760 tatagcgtct tatcttgctg atataacaac agctaaaccg tatgcaccag gcaaatttga 14820 gaaaaagaca acttcagaag cattgaaaga tgtgttgagc gatacaggtt gggaagtttc 14880 tgaacaaacc gaatacgatg gcttacgtac tacgtcatgg acttcttatc aaactagata 14940 tgaagtttta aagcaattat gtacaaccta taaaatggtt ttagattttt atattgagct 15000 tagctctaat accgtcaaag gtagatatgt agtactcaaa aagaaaaaca gcttattcaa 15060 aggtaaagaa attgaatatg gtaaagattt agtcgggtta actaggaaga ttgatatgtc 15120 agaaatcaaa acagcattaa ttgctgtggg acctgaaaat gacaaaggga agcgtttaga 15180 gctagttgtg acagatgacg aagcgcaaag tcaattcaac ctacctatgc gctatatttg 15240 ggggatatat gaaccacaat cagatgatca aaatatgaat gaaacacgat taagttcttt 15300 agccaaaaca gagttaaata aacgtaagtc ggcagttatg tcatatgaga ttacttctac 15360 tgatttggaa gttacgtatc cgcacgagat tatatcaatt ggcgatacag tcagagtaaa 15420 acatagagat tttaacccgc cattgtatgt agaggcagaa gttattgctg aagaatataa 15480 cataatttca gaaaatagca catatacatt cggtcaacct aaagagttca aagaatcaga 15540 attacgagaa gagtttaaca agcgattgaa cataatacat caaaagttaa acgataatat 15600 tagcaatatc aacactatag ttaaagatgt tgtagatggt gaattagaat actttgaacg 15660 caaaatacac aaaagtgata caccgccaga aaatccagtc aatgatatgc tttggtatga 15720 tacaagtaac cctgatgttg ctgtcttgcg tagatattgg aatggtcgat ggattgaagc 15780 aacaccaaat gatgttgaaa aattaggtgg tataacaaga gagaaagcgc tattcagtga 15840 attaaacaat atttttatta atttatctat acaacacgct agtcttttgt cagaagctac 15900 agaattactg aatagcgagt acttagtaga taatgatttg aaagcggact tacaagcaag 15960 tttagacgct gtgattgatg tttataatca aattaaaaat aatttagaat ctatgacacc 16020 cgaaactgca acgattggtc ggttggtaga tacacaagct ttatttcttg agtatagaaa 16080 gaaattacaa gatgtttata cagatgtaga agatgtcaaa atcgccattt cagatagatt 16140 taaattatta cagtcacaat acactgatga aaaatataaa gaagcgttgg aaataatagc 16200 aacaaaattt ggtttaacgg tgaatgaaga tttgcagtta gtcggagaac ctaatgttgt 16260 taaatcagct attgaagcag ctagagaatc cacaaaagaa caattacgtg actatgtaaa 16320 aacatcggac tataaaacag acaaagacgg tattgttgaa cgtttagata ctgctgaagc 16380 tgagagaacg actttaaaag gtgaaatcaa agataaagtt acgttaaacg aatatcgaaa 16440 cggattggaa gaacaaaaac aatatactga tgaccagtta agtgatttgt ccaataatcc 16500 tgagattaaa gcaagtattg aacaagcaaa tcaagaagcg caagaagctt taaaatcata 16560 cattgatgct caagatgatc ttaaagagaa ggaatcgcaa gcgtatgctg atggtaaaat 16620 ttcggaagaa gagcaacgcg ctatacaaga tgctcaagct aaacttgaag aggcaaaaca 16680 aaacgcagaa ctaaaggcta gaaacgctga aaagaaagct aatgcttata cagacaacaa 16740 ggtcaaagaa agcacagatg cacagaggaa aacattgact cgctatggtt ctcaaattat 16800 acaaaatggt aaggaaatca aattaagaac tactaaagaa gagtttaatg caaccaatcg 16860 tacactttca aatatattaa acgagattgt tcaaaatgtt acagatggaa caacaatcag 16920 atatgatgat aacggagtgg ctcaagcttt gaatgtgggg ccacgtggta ttagattaaa 16980 tgctgataaa attgatatta acggtaatag agaaataaac cttcttatcc aaaatatgcg 17040 agataaagta gataaaaccg atattgtcaa cagtcttaat ttatcaagag agggtcttga 17100 tatcaatgtt aatagaattg gaattaaagg cggtgacaat aacagatatg ttcaaataca 17160 gaatgattct attgaactag gtggtattgt gcaacgtact tggagaggga aacgttcaac 17220 agacgatatt tttacgcgac tgaaagacgg tcacctaaga tttagaaata acaccgctgg 17280 cggttcactt tatatgtcac attttggtat ttcgacttat attgatggtg aaggtgaaga 17340 cggtggttca tctggtacga ttcaatggtg ggataaaact tacagtgata gtggcatgaa 17400 tggtataaca atcaattcct atggtggtgt cgttgcacta acgtcagata ataatcgggt 17460 tgttctggag tcttacgctt catcgaatat caaaagcaaa caggcaccgg tgtatttata 17520 tccaaacaca gacaaagtgc ctggattaaa ccgatttgca ttcacgctgt ctaatgcaga 17580 taatgcttat tcgagtgacg gttatattat gtttggttct gatgagaact atgattacgg 17640 tgcgggtatc aggttttcta aagaaagaaa taaaggtctt gttcaaattg ttaatggacg 17700 atatgcaaca ggtggagata caacaatcga agcagggtat ggcaaattta atatgctgaa 17760 acgacgtgat ggtaataggt atattcatat acagagtaca gacctactgt ctgtaggttc 17820 agatgatgca ggagatagga tagcttctaa ctcaatttat agacgtactt attcggccgc 17880 agctaatttg catattactt ctgctggcac aattgggcgt tcgacatcag cgcgtaaata 17940 caagttatct atcgaaaatc aatataacga tagagatgaa caactggaac attcaaaagc 18000 tattcttaac ttacctatta gaacgtggtt tgataaagct gagtctgaaa ttttagctag 18060 agagctgaga gaagatagaa aattatcgga agacacctat aaacttgata gatacgtagg 18120 tttgattgct gaagaggtgg agaatttagg attaaaagag tttgtcacgt atgatgacaa 18180 aggagaaatt gaaggtatag cgtatgatcg tctatggatt catcttatcc ctgttatcaa 18240 agaacaacaa ctaagaatca agaaattgga ggagtcaaag aatgcaggat aacaaacaag 18300 gattacaagc taatcctgaa tatacaattc attatttatc acaggaaatt atgaggttaa 18360 cacaagaaaa cgcgatgtta aaagcgtata tacaagaaaa taaagaaaat caacaatgtg 18420 ctgaggaaga gtaatcctta gcactatttt tatacaaaaa tttaaggagg tcatttaatt 18480 atggcaaaag aaattatcaa caatacagaa aggtttattt tagtacaaat cgacaaagaa 18540 ggtacagaac gtgtagtata tcaagatttc acaggaagtt ttacaacttc tgaaatggtt 18600 aaccatgctc aagattttaa atctgaagaa aacgctaaga aaattgcgga gacgttaaat 18660 ttgttatatc aattaactaa caaaaaacaa cgtgtgaaag tagttaaaga agtagttgaa 18720 agatcagatt tatctccaga ggtaacagtt aacactgaaa cagtatgaaa agctatgagt 18780 tagatactca tagtctttat tcttttagaa agcgggtgta ctgaattggg gtggttcaaa 18840 aaacacgaac atgaatggcg catcagaagg ttagaagaga atgataaaac aatgctcagc 18900 acactcaacg aaattaaatt aggtcaaaaa acccaagagc aagttaacat taaattagat 18960 aaaaccttag atgctattca aaaagaaaga gaaatagatg aaaagaataa gaaagaaaat 19020 gataagaaca tacgtgatat gaaaatgtgg gtgcttggtt tagttgggac aatatttggg 19080 tcgctaatta tagcattatt gcgtatgctt atgggcatat aagagaggtg attaccatgt 19140 tcggattaaa ttttggagct tcgctgtgga cgtgtttctg gtttggtaag tgtaagtaat 19200 agttaagagt cagtgcttcg gcactggctt tttattttgg ataaaaggag caaacaaatg 19260 gatgcaaaag taataacaag atacatcgta ttgatcttag cattagtaaa tcaattctta 19320 gcgaacaaag gtattagccc aattccagta gacgatgaaa ctatatcatc aataatactt 19380 actgtagtcg ctttatatac aacgtataaa gacaatccaa catctcaaga aggtaaatgg 19440 gcaaatcaaa aattaaagaa atataaagct gaaaataagt atagaaaagc aacagggcaa 19500 gcgccaatta aagaagtaat gacacctacg aatatgaacg acacaaatga tttagggtag 19560 gtggttgata tatgttaatg acaaaaaatc aagcagaaaa atggtttgac aattcattag 19620 ggaaacaatt caacccagat ggttggtatg gatttcagtg ttatgattac gccaatatgt 19680 tctttatgtt agcgacaggc gaaaggctgc aaggtttata tgcttataat atcccgtttg 19740 ataataaagc aaagattgaa aaatatggtc aaataattaa aaactatgac agctttttac 19800 cgcaaaagtt ggatattgtc gttttcccgt caaagtatgg tggcggagct ggacacgttg 19860 aaattgttga gagcgcaaat ttaaatactt tcacatcatt tggtcaaaac tggaacggta 19920 aaggttggac taatggcgtt gcgcaacctg gttggggtcc tgaaactgtg acaagacatg 19980 ttcattatta tgacaatcca atgtatttta ttaggttaaa cttccctaac aacttaagcg 20040 ttggcaataa agctaaaggt attattaagc aagcgactac aaaaaaagag gcagtaatta 20100 aacctaaaaa aattatgctt gtagccggtc atggttataa cgatcctgga gcagtaggaa 20160 acggaacaaa cgaacgcgat tttatacgta aatatataac gcctaatatc gctaagtatt 20220 taagacatgc aggacatgaa gttgcattat acggtggctc aagtcaatca caagatatgt 20280 atcaagatac tgcatacggt gttaatgtag gcaataaaaa agattatggc ttatattggg 20340 ttaaatcaca ggggtatgac attgttctag aaatacattt agacgcagca ggagaaagcg 20400 caagtggtgg gcatgttatt atctcaagtc aattcaatgc agatactatt gataaaagta 20460 tacaagatgt tattaaaaat aacttaggac aaataagagg tgtgacacct cgtaatgatt 20520 tactaaatgt taatgtatca gcagaaataa atataaatta tcgtttatct gaattaggtt 20580 ttattactaa taaaaatgat atggattgga ttaagaaaaa ctatgacttg tattctaaat 20640 taatagccgg tgcgattcat ggtaagccta taggtggttt ggtagctggt aatgttaaaa 20700 catcagctaa aaacaaaaaa aatccaccag tgccagcagg ttatacactc gataagaata 20760 atgtccctta taaaaaagaa caaggcaatt acacagtagc taatgttaaa ggtaataatg 20820 taagagacgg ttattcaact aattcaagaa ttacaggggt attacccaac aacacaacaa 20880 ttacgtatga cggtgcatat tgtattaatg gttatagatg gattacttat attgctaata 20940 gtggacaacg tcgttatata gcgacaggag aggtagacaa ggcaggtaat agaataagta 21000 gttttggtaa gtttagcacg atttagtatt tacttagaat aaaaattttg ctacattaat 21060 tatagggaat cttacagtta ttaaataact atttggatgg atgttaatat tcctatacac 21120 tttttaacat ttctctcaag atttaaatgt agataacagg caggtacttc ggtacttgcc 21180 tattttttta tgttatagct agccttcggg ctagtttttt gttatgatgt gttacacatg 21240 catcaactat ttacatctat ccttgttcac ccaagcatgt cactggatgt tttttcttgc 21300 gatagagagc atagttttca tactactccc cgtagtatat atgactttag cattcccgta 21360 taacagttta cggggtgctt ttatgttata attgctttta tatagtagga gtgaactata 21420 tagccgggca gaggccatgt atctgactgt tggtcccaca ggagacatct tccttgtcat 21480 cactcgatac atatatctta acaacataga aatgttacat tcgctataac cgtatcttaa 21540 tcgatacggt tatatttatt cccctacaac caacaaaacc acagatccta ttaatttagg 21600 attgtggtta ttttttgcgt ttttttgggg caaaaaaagg gcagattatt tgaaaaaggg 21660 caaacgcttg tggaaaagct aaaaggttaa aaatgacaaa aaccttgata caacagtgtt 21720 tttggacgct cgtgtacgtt agagaatgac cggtttacca tcatacaagg gtgggattaa 21780 cttgtgttaa aaagccttta atatcagttg ttacaaagga tttgtagcgt ctttaaaaat 21840 aaaaaagggc agaaaaaggg cagatacctt ttagtacaca agtttttcta atttttgctc 21900 taactctctg tccattttct ctgttacatg tgtatacacc tttatagtcg ttttttcatc 21960 tgtatgtcct actcttttca taattgcttt taacgatata ttcatttccg ccaataaact 22020 tatgtgtgta tgccttagtg tgtgagtagt aactttttta tttatattta atgattctgc 22080 agctgaggac aatcgtttgt ttatcctact gccttgcata ggatttcctt ggcaagttgt 22140 gaatataaac cctctatcaa catagcttgg ttcccattgt tgcatctttt tattttctaa 22200 cattattttt ttcaatacat ttgctatcct tgaattgatg gcgatttttc ttcttgaacc 22260 tgcggtctta gtagtatctt tgtgaccaaa tccagcatta catttgattc tgtgaatagt 22320 gccattaata gcgatcgttt tatttttgag gtcaacatct ttaacttgga gagctaataa 22380 ctcacctatg cgcatacctg ttaaagcttg aacttctaca gccccagcaa ctaaaatacg 22440 agctctatac tgcatgttat tatcgttcag tataaaatcg cgtatctgta ttacctgttc 22500 catctctaaa tagttataca ttttcgcttc ttctttttct atatcttcta tcgtcttact 22560 cttctttggt agtgtgacgc tatttaatat gtgttcgttt ggataattgt aaaatttaac 22620 ggcgtattta atagcttctt tcatatgtcc aagttgacgc tttacctgat ttgcagaata 22680 tacgtttgat aatttgttaa taaatgtttg catgtacttt gtatcaattt tgtttaaaag 22740 taaattttga gaactgttct ttttgatgtt tttgattctt gttttcaaat tatcaagcgt 22800 cgttacttta aagccagatg tttttatatg atattcaagc cattcatcta ataacgcgtg 22860 aaaagtcaaa gtttttaatt cgcttgacga cttgttgttt agtttttctt ttattttttc 22920 ttctaaacga aacattgcct ctttttgcga ttgctttgta ttcttattca agacaacact 22980 tacacgtttc catttatctg tatacggatc tttgtatttc tcgtagtatc tatacttcgt 23040 ttcattgttc ttatttttaa atttttcaaa ccacatttta catccctcct caaaattggc 23100 aaaaaataat aagggtaggc gggctaccca tgaaaattgt ataaaaaaag acgcctgtat 23160 aaaatacaga cgccacttat aattataaga ttacatggtt aattaccaaa aatggtaacg 23220 aatatatacg tgttttaaag gataaacctt taatatatta aaattatatc atcttatatc 23280 agggatctgc aatatattat tattaattct atttatcagt aacataatat ccgaagaatc 23340 tattactgga tttttaattt tttggggtaa aacttttctt atgcgaaact tactaatcgg 23400 ctggaaagaa tttatgcaag cgtaactatt accttttaat ttttttacct tatcaattgc 23460 tgatactatg ttattaatgt ttctgtcaat tttatttaat ttattttcaa tttctaaact 23520 atcagatata aattcaataa aataatcttt agtgatgaat tctgtgttgt ttttttggta 23580 ttttttatcg aaaacttctt ttaatatagc tgaattattt tgcgcgctaa ttaaatttaa 23640 aaacaatctt aaataatact cccatttcaa atcaaaattc atctttaaat actttttgtt 23700 ttctttagag gataagggaa taacatttac tatatcctcc gtattagaat catttttatt 23760 catcactatt gcaaagtgtg aattagaaaa ttctttatta acgtttatac cgaaatctac 23820 aaaaactatt tctccttgtt taaactttgg ataaaaacct ttatggtttt tttcaccttc 23880 aaatctcttg agtaaatagt gaatatctga atctaacttt ttaaattttg gatttccaga 23940 agtttttaat ttattaatgc gtttttctat attatgcgtc atcatttctc ctttattctc 24000 gctcacactc tcaccaccat tcaacgtcta cacttgtagg cgttttttga ttagtaaaat 24060 cataatgaat cttctttggt taacttatcg ccatctattt tttgtgaaat aaattccaag 24120 tatttacgcg cattatgtga cgataaatct ttaggtaact cataagtgaa tggttgatta 24180 ccactagtta aaacttcata tactatagtt tcttttttta ttttgcaatt agttattttc 24240 attataaact ccttttaaac actgctgaaa tagacgtctt tttcaaataa gcatgattaa 24300 tactttaatt ctttaatcca catatattta aaagtgaggt agtaggtaat aaatataaga 24360 cttaaagtta agattgcttt tttcatgtca atttctcctt tgtttatatt tatattaaag 24420 cgctaaatat acgttattaa tcacaataca actttgccca ttactttaat atcactaaac 24480 gaagcgactt tgatatcatc atacttcgga tttagagata ccaaattaat atagtcttcg 24540 catatatcta cacgcttgat aagacttact ccatctaata caacgagtgc aattgtacca 24600 tctttaatag aatcttcttt cttaataaaa gcgtatgttc cttgttttaa cataggttcc 24660 attgaatcac cattaactaa aatacaaaaa tcagcatttg atggcgtttc gtcttcttta 24720 aaaaatactt cttcatgcaa tatgtcatca tataattctt ctcctatgcc agcaccagtt 24780 gcaccacatg caatatacga tactagttta gactctttat attcatctat agaagtgact 24840 ttattctgtt catctaattg ctcatttgca tagttaagta cgttttcttg gcggggaggt 24900 gtgagttgag aaaatatgtt attgattttt gacattatcg tttcatcttg acgttcttcg 24960 tcaggaactc gataagaatc tacatcatac cccataagcc acgcttcacc gacatttaaa 25020 gttttagata ataagaataa tttatgttgg tctggagaag accttccatt aacatactgg 25080 gataagtgac tttttgacat tttaatattc aattcttttt gaaagggttt cgacttttct 25140 agaatatcta cttgacgcaa gttcctatct ttcataattt gttttaatct ttcagaagtg 25200 ttttgcattg gtaatgcctc cttgaaattc attatatagg aagggaaata aaaatcaata 25260 caaaagttca acttttttaa ctttttgtgt tgacattgtt caaaattggg gttatagtta 25320 ttatagttca aatgtttgaa cttaggaggt gattatttga atactaatac aacttttgat 25380 ttttcgttat tgaacggtaa gatagtcgaa gtgtactcga cacaatttaa ctttgctata 25440 gctttaggtg tatcagaaag aactttgtct ttgaagttga acaacaaagt accatggaaa 25500 acaacagaca ttattaaagc ttgtaagtta ttgggaatac ctataaaaga tgttcacaaa 25560 tattttttta aacagaaagt tcaaatgttt gaacttaata agtaaaggag gcataacaca 25620 tgcaagaacg agaaaaggtt aataaaagta acacatcttc aaatgaagca tcaaaacctt 25680 ttaggacaaa ttgaagctta cgacaaaacg cttaaagaaa taaagtacac tcgagacctt 25740 tacaacaaac acctaagcat gaacaacgaa gacgcattcg ctggtttgga aatggtagag 25800 gatgaaatta ctaaaaagct acgaagtgct atcaaagagt tccaaaaagt agtgaaagcg 25860 ttagacaagc ttaacggtgt tgaaagcgat aacaaagtta ctgatttaac agagtggcgg 25920 aaagtgaatc agtaacattc acttcttaat ataaccacgc ttatcaacat ccacattgag 25980 cagatgtgag cgagagctgg cgatgatatg agccgcgttt aaatacattc gatagtcatt 26040 gcgataaccg tctgctgaat gtgggtgttg aggaaaaagg aggatactca aatgcaagca 26100 ttacaaacat ttaattttaa agagctacca gtaagaacag tagaaattga aaacgaacct 26160 tattttgtag gaaaagatat tgctgagatt ttaggatatg caagatcaaa caatgccatt 26220 agaaatcatg ttgatagcga ggacaagctg acgcaccaat ttagtgcatc aggtcaaaac 26280 agaaatatga tcattatcaa cgaatcagga ttatacagtc taatcttcga tgcttctaaa 26340 caaagcaaaa acgaaaaaat tagagaaacc gctagaaaat tcaaacgctg ggtaacatca 26400 gatgtcctac cagctattcg caaacacggt atatacgcaa cagacaatgt aattgaacaa 26460 acattaaaag atccagacta catcattaca gtgttgactg agtataagaa agaaaaagag 26520 caaaacttac ttttacaaca gcaagtagaa gttaacaaac caaaagtatt attcgctgac 26580 tcggtagctg gtagtgataa ttcaatactt gttggagaac tagcgaaaat acttaaacaa 26640 aacggtgttg atataggaca aaacagattg ttcaaatggt taagaaataa tggatatctc 26700 attaaaaaga gtggagaaag ttataactta ccaactcaaa agagtatgga tctaaaaatc 26760 ttggatatca aaaaacgaat aattaataat ccagatggtt caagtaaagt atcacgtaca 26820 ccaaaagtaa caggcaaagg acaacaatac tttgttaata agtttttagg agaaaaacaa 26880 acatcttaaa aggaggaaca caatggaaca aatcacatta accaaagaag agttgaaaga 26940 aattatagca aaagaagtta gagaggctat aaatggcaag aaaccaatca gttcaggttc 27000 aattttcagt aaagtaagaa tcaataatga cgatttagaa gaaatcaata aaaaactcaa 27060 tttcgcaaaa gatttgtcgc taggaagatt gaggaagctc aatcatccga ttccgctaaa 27120 aaagtatcag catggcttcg aatcaattca tcaaaaagct tatgtacaag atgttcatga 27180 ccatattaga aaattaacat tatcaatttt tggagtgaca cttaattcag acttgagtga 27240 aagtgaatac aacctagcag caaaagttta tcgagaaatc aaaaactatt atttatacat 27300 ctatgaaaag agagtttcag aattaactat cgatgatttc gaataaagga ggaacaacaa 27360 atgttacaaa aatttagaat tgcgaaagaa aaaaataaat taaaactcaa attactcaag 27420 catgctagtt actgtttaga aagaaacaac aaccctgaac tgttgcgagc agttgcagag 27480 ttgttgaaaa aggttagcta aattcaacgg taaggatttg ccctgcctcc acacttagag 27540 tttgagatcc aacaaacaca taagttttag tagggtctag aaaaaatgtt tcgatttcct 27600 cttttgtaac agtttcaatt ccttcatatc ctggaaaaac aattttcttt aaatccgaaa 27660 catgtttttt tgaaccatcc tttaaagtaa ctagaagttt catacttatc acctccttag 27720 gttgataaca acattataca cgaaaggagc ataaacaata tgcaagcatt acaaacaaat 27780 tcgaacatcg gagaaatgtt caatattcaa gaaaaagaaa atggagaaat cgcaatcagc 27840 ggtcgagaac ttcatcaagc attagaagtt aagacagcat ataaagattg gtttccaaga 27900 atgcttaaat acggatttga agaaaataca gattacacag ctatcgctca aaaaagagca 27960 acagctcaag gcaatatgac tcactatatt gaccacgcac tcacactaga cactgcaaaa 28020 gaaatcgcaa tgattcaacg tagtgaacct ggcaaacgtg caagacaata tttcatccaa 28080 gttgaaaaag catggaacag cccagaaatg attatgcaac gtgctttaaa aattgctaac 28140 aacacaatca atcaattaga aacaaagatt gcacgtgaca aaccaaaaat tgtatttgca 28200 gatgcagtag ctactactaa gacatcaatt ttagttggag agttagcaaa gatcattaaa 28260 caaaacggta taaacatcgg gcaacgcaga ttgtttgagt ggttacgtca aaacggattc 28320 cttattaaac gcaagggtgt ggattataac atgcctacac agtattcaat ggaacgtgag 28380 ttattcgaaa ttaaagaaac atcaatcaca cattcggacg gtcacacatc aattagtaag 28440 acgccaaaag taacaggtaa aggacaacaa tactttgtta acaagttttt aggagaaaaa 28500 caaacaactt aataggagga attacaaatg aacgcactat acaaaacaac cctcctcatc 28560 acaatggcag ttgtgacgtg gaaggtttgg aagattgaga agcacactag aaaacctgtg 28620 attagtagca gggcgttgag tgactatcta aacaacaaat ctttaaccat accgaaagat 28680 gctgaaaatt ctactgaatc tgctcgtcgc cttttgaagt tcgccgaaca aactattagc 28740 aaataacaac attatacacg aaaggaaaga tagaaatgcc aaaaatcata gtaccaccaa 28800 caccagaaaa cacatataga ggcgaagaaa aatttgtgaa aaagttatac gcaacaccta 28860 cacaaatcca tcaattgttt ggagtatgta gaagtacagt atacaactgg ttgaaatatt 28920 accgcaaaga taatttaggt gtagaaaatt tatacattga ttattcacca acaggcactc 28980 tgattaatat ttctaaattg gaagagtatt tgatcagaaa gcataaaaaa tggtattagg 29040 aggatattaa atgagcaaca tttataaaag ctacctagta gcagtattat gcttcacagt 29100 cttagcgatt gtacttatgc cgtttctata cttcactaca gcatggtcaa ttgcgggatt 29160 cgcaagtatc gcaacattca tgtactacaa agaatgcttt ttcaaagaat aaaaaaactg 29220 ctacttgttg gagcaagtaa cagtatcaaa cacttaagaa aaaattcatg ttcaatataa 29280 aacgaaaaac ggaggaagtc aagatgtatt acgaaatagg cgaaatcata cgcaaaaata 29340 ttcatgttaa cggattcgat tttaagctat tcattttaaa aggtcatatg ggcatatcaa 29400 tacaagttaa agatatgaac aacgtaccaa ttaaacatgc ttatgtcgta gatgagaatg 29460 acttagatat ggcatcagac ttatttaacc aagcaataga tgaatggatt gaagagaaca 29520 cagacgaaca ggacagacta attaacttag tcatgaaatg gtaggaggtc gctatgaagc 29580 agactgtaac ttatatcatt cgtcataggg atatgccaat ttatataact aacaaaccaa 29640 ctgataacaa ttcagatatt agttactcca caaatagaaa tagagctagg gagtttaacg 29700 gtatggaaga agcgagtatc aatatggatt atcacaaagc aatcaagaaa acagtgacag 29760 aaactattga gtacgaggag gtagaacatg actgaggaaa aacaagaacc acaagaaaaa 29820 gtaagcatac tcaaaaaact aaagataaat aatatcgctg agaaaaataa aaggaaattc 29880 tataaatttg cagtatacgg aaaaattggc tcaggaaaaa ccacgtttgc tacaagagat 29940 aaagacgctt tcgtcattga cattaacgaa ggtggaacaa cggttactga cgaaggatca 30000 gacgtagaaa tcgagaacta tcaacacttt gtttatgttg taaatttttt acctcaaatt 30060 ttacaggaga tgagagaaaa cggacaagaa atcaatgttg tagttattga aactattcaa 30120 aaacttagag atatgacatt gaatgatgtg atgaaaaata agtctaaaaa accaacgttt 30180 aatgattggg gagaagttgc tgaacgaatt gtcagtatgt acagattaat aggaaaactt 30240 caagaagaat acaaattcca ctttgttatt acaggtcatg aaggtatcaa caaagataaa 30300 gatgatgaag gtagcactat caaccctact atcactattg aagcgcaaga acaaattaaa 30360 aaagctatta cttctcaaag tgatgtgtta gctagggcaa tgattgaaga atttgatgat 30420 aacggagaaa agaaagctag atatattcta aacgctgaac cttctaatac gtttgaaaca 30480 aagattagac attcaccttc aataacaatt aacaataaga aatttgcaaa tcctagcatt 30540 acggacgtag tagaagcaat tagaaatgga aactaaaaat taattaaaag gacggtattt 30600 aattatgaaa atcacaggac aagcgcaatt tactaaagaa acaaatcaag aaaagtttta 30660 taacggctca gcagggtttc aagctggaga attcacagtg aaagttaaaa atattgaatt 30720 caatgataga gaaaatagat atttcacaat cgtatttgaa aatgatgaag gcaaacaata 30780 taaacataat caatttgtac cgccgtataa atatgatttc caagaaaaac aattgattga 30840 attagttact cgattaggta ttaagttaaa tcttcctagc ttagattttg ataccaatga 30900 tcttattggt aagttttgtc acttggtatt gaaatggaaa ttcaatgaag atgaaggtaa 30960 gtattttacg gatttttcat ttattaaacc ttacaaaaag ggcgatgatg ttgttaacaa 31020 acctattccg aagacagata agcaaaaagc tgaagaaaat aacggggcac aacaacaaac 31080 atcaatgtct caacaaagca atccatttga aagcagtggc caatttggat atgacgacca 31140 agatttagcg ttttaaggtg tggtttaaat gcaatacatt acaagatacc agaaagataa 31200 cgacggtact tattccgtcg ttgctactgg tgttgaactt gaacaaagtc acattgactt 31260 actagaaaac ggatatccac taaaagcaga agtagaggtt ccggacaata aaaaactatc 31320 tatagaacaa cgcaaaaaaa tattcgcaat gtgtagagat atagaacttc actggggcga 31380 accagtagaa tcaactagaa aattattaca aacagaattg gaaattatga aaggttatga 31440 agaaatcagt ctgcgcgact gttctatgaa agttgcaagg gagttaatag aactgattat 31500 agcgtttatg tttcatcatc aaatacctat gagtgtagaa acgagtaagt tgttaagcga 31560 agataaagcg ttattatatt gggctacaat caaccgcaac tgtgtaatat gcggaaagcc 31620 tcacgcagac ctggcacatt atgaagcagt cggcagaggc atgaacagaa acaaaatgaa 31680 ccactatgac aaacatgtat tagcgttatg tcgcgaacat cacaacgagc aacatgcgat 31740 tggcgttaag tcgtttgatg ataaatacca cttgcatgac tcgtggataa aagttgatga 31800 gaggctcaat aaaatgttga aaggagagaa aaaggaatga atagactaag aataataaaa 31860 atagcactcc taatcgtcat cttggcggaa gagattagaa atgctatgca tgctgtaaaa 31920 gtggagaaaa ttttaaaatc tccgtttagt taatacaggt ttttacaaaa gctttaccat 31980 aggcggacaa actaattgag ccttttttga tgtctattac ccaggggctg taatgtaact 32040 ttaatacttc aaattcaatg ccagaaagtt tacttattgt ttctaggttg tgtcctgact 32100 ttaacattct tttaacaaat tctaatcccg aaacaaatct ttgtttttct ataatcttat 32160 taaagtgatt taaaaactga ggagcataaa acttattata aattcctttt tttgttaagt 32220 aagacatgtc aaaagtttca tttaaaaccc ctaaccttac taggttatta attgaaattt 32280 cggttgattc tatatctaac ggagagtctt ttattaacgt gtccgatata ttcataccgt 32340 cattctttgg gtttaaaacc gctctatatt taacggcagg atgtacttcg tgattcttta 32400 aatgttttaa aagaatagca tcatttgggg ataattgttt aattatttca acaaatgaat 32460 ggtgggttaa tgagtttttt ctgtcatcca tagatgatgc tattagtttt gcgaacatat 32520 tacttaaagt tttttcacta atgtaaaact ttgaagcttc tagagcagga cctagaagag 32580 aaaattgtgg ttcttgtaaa ttatttttag gtacagaaga tatttctttt ttaaattgtt 32640 ctttgaattt ttcaaattct acttctcttt gataaataac tttatccaca taaaggtgga 32700 atttcccaaa gacaagttcc caagttttag agaatgtttc tacaggccct tttgatgcgc 32760 cttcaataat tttatcaata cctttaccta aaataggatc cataattatt cacccccaat 32820 ctaacgcaat agcgataata aaattatacc agaaaggaga atcaacatga ctgaccaacc 32880 aagttactac tcaataatta cagcaaatgt cagatacgat aaccgactta ctgacagcga 32940 aaagttactt tttgcagaaa taacatcttt aagtaacaaa tacggatact gcacagcaag 33000 taatggttac tttgcaactt tatacaacgt tgttaaggaa actatatctc gtagaatttc 33060 gaaccttacc aactttggtt atctaaaaat cgaaattatc aaagaaggta atgaagttaa 33120 acaaaggaag atgtacccct tgacgcaaac gtcaatacct attgacgcaa aaatcaatac 33180 ccctattgat aattctgtca atacccctat tgacgcaaat gtcaaagaga atattacaag 33240 tattaataat acaagtaata acaatataaa tagaatagat atattgtcgg gcaacccgac 33300 agcatcttct ataccctata aagaaattat cgattactta aacaaaaaag cgggcaagca 33360 ttttaaacac aatacagcta aaacaaaaga ttttattaaa gcaagatgga atcaagattt 33420 taggttggag gattttaaaa aggtgattga tatcaaaaca gctgagtggc taaacacgga 33480 tagcgataaa taccttagac cagaaacact ttttggcagt aaatttgagg ggtacctcaa 33540 tcaaaaaata caaccaactg gcacggatca attggaacgc atgaagtacg acgaaagtta 33600 ttgggattag ggggatatta tgaaaccact attcagcgaa aagataaacg aaagcttgaa 33660 aaaatatcaa cctactcatg tcgaaaaagg attgaaatgt gagagatgtg gaagtgaata 33720 cgacttatat aagtttgctc ctactaaaaa acacccgaat ggttacgagt ataaagacgg 33780 ttgcaaatgt gaaatctatg aggaatataa gcgaaacaag caacggaaga taaacaacat 33840 attcaatcaa tcaaacgtta atccgtcttt aagagatgca acagtcaaaa actacaagcc 33900 acaaaatgaa aaacaagtac acgctaaaca aacagcaata gagtacgtac aaggcttctc 33960 tacaaaagaa ccaaaatcat taatattgca aggttcatac ggaactggta aaagccacct 34020 agcatacgct atcgcaaaag cagtcaaagc taaagggcat acggttgctt ttatgcacat 34080 accaatgttg atggatcgta tcaaagcgac atacaacaaa aatgcagtag agactacaga 34140 cgagctagtc agattgctaa gtgatattga tttacttgta ctagatgata tgggtgtaga 34200 aaacacagag cacactttaa ataaactttt cagcattgtt gataacagag taggtaaaaa 34260 caacatcttt acaactaact ttagtgataa agaactaaat caaaatatga actggcaacg 34320 tataaattcg agaatgaaaa aaagagcaag aaaagtaaga gtaatcggag acgatttcag 34380 ggagcgagat gcatggtaac caaagaattt ttaaaaacta aacttgagtg ttcagatatg 34440 tacgctcaga aactcataga tgaggcacag ggcgatgaaa ataggttgta cgacctattt 34500 atccaaaaac ttgcagaacg tcatacacgc cccgctatcg tcgaatatta aggagtgtta 34560 aaaatgccga aagaaaaata ttacttatac cgagaagatg gcacagaaga tattaaggtc 34620 atcaagtata aagacaacgt aaatgaggtt tattcgctca caggagccca tttcagcgac 34680 gaaaagaaaa ttatgactga tagtgaccta aaacgattca aaggcgctca cgggcttcta 34740 tatgagcaag aattaggttt acaagcaacg atatttgata tttagaggtg gacgatgagt 34800 aaatacaacg ctaagaaagt tgagtacaaa ggaattgtat ttgatagcaa agtagagtgt 34860 gaatattacc aatatttaga aagtaatatg aatggcacta attatgatca tatcgaaata 34920 caaccgaaat tcgaattatt accaaaacta gataaacaac gaaagattga atatattgca 34980 gacttcgcgt tatatctcga tggcaaactg attgaagtta tcgacattaa aggtatgcca 35040 accgaagtag caaaacttaa agctaagatt ttcagacata aatacagaaa cataaaactc 35100 aattggatat gtaaagcgcc taagtataca ggtaaaacat ggattacgta cgaggaatta 35160 attaaagcaa gacgagaacg caaaagagaa atgaagtgat ctaatgcaac aacaagcata 35220 tataaatgca acgattgata taaggatacc tacagaagtt gaatatcagc attttgatga 35280 tgtggataaa gaaaaagaag cgctggcaga ttacttatat aacaatcctg acgaaatact 35340 agagtatgac aatttaaaaa ttagaaacgt aaatgtagag gtggaataaa tgggcagtgt 35400 tgtaatcatt aataataaac catataaatt taacaatttt gaaaaaagaa ataatggcaa 35460 agcgtgggat aaatgctgga attgtttcta aacgtgttag aggttgttgg gagttttcag 35520 aagctttaga cgcgccttat ggcatgcacc taaaagaata tagagaaatg aaacaaatgg 35580 aaaagattaa acaagcgaga ctcgaacgtg aattggaaag agagcgaaag aaagaggctg 35640 agctacgtaa gaagaagcca catttgttta atgtacctca aaaacattca cgtgatccgt 35700 actggttcga tgtcacttat aaccaaatgt tcaagaaatg gagtgaagca taatgagcat 35760 aatcagtaac agaaaagtag atatgaacaa aacgcaagac aacgttaagc aacctgcgca 35820 ttacacatac ggcgacattg aaattataga ttttattgaa caagttacgg cacagtaccc 35880 accacaatta gcattcgcaa taggtaatgc aattaaatac ttgtctagag caccgttaaa 35940 gaatggtcat gaggatttag caaaggcgaa gttttacgtc gatagagtat ttgacttgtg 36000 ggagtgatga ccatgacaga tagcggacgt aaagaatact taaaacattt tttcggctct 36060 aagagatatc tgtatcagga taacgaacga gtggcacata tccatgtagt aaatggcact 36120 tattactttc acggtcatat cgtgccaggt tggcaaggtg tgaaaaagac atttgataca 36180 gcggaagagc ttgaaacata tataaagcaa agtgatttgg aatatgagga acagaagcaa 36240 ctaactttat tttaaaaggg cggaaacaat gaaaatcaaa attgaaaaag aaatgaattt 36300 acctgaactt atccaatggg cttgggataa ccccaagtta tcaggtaata aaagattcta 36360 ttcaaatgat gttgagcgca actgttttgt gacttttcat gttgatagca tcttatgtaa 36420 tgtgactgga tatgtatcaa ttaacgataa atttactgtt caagaggaga tataacaatg 36480 aaaatcaaag ttaaaaaaga aatgagatta gatgaattaa ttaaatgggc gcgagaaaat 36540 ccggatctat cacaaggaaa aatatttttt tcaacaggat ttagtgatgg attcgttcgt 36600 tttcatccaa atacaaataa gtgttcgacg tcaagtttta ttccaattga tatccccttc 36660 atagttgata ttgaaaaaga agtaacggaa gagactaagg ttgataggtt gattgaatta 36720 ttcgagattc aagaaggaga ctataactct acactatatg agaacactag tataaaagaa 36780 tgtttatatg gcagatgtgt gcctaccaaa gcattctaca tcttaaacga tgacctaact 36840 atgacgttaa tctggaaaga tggggagttg ctagtatgat gttgaaattt aaagcttggg 36900 ataaagataa aaaagttatg agtattattg acgaaatcga ttttaatagt gggtacattt 36960 tgatttcaac aggttataaa agtttcaatg aagtaaaact attacaatac acaggattta 37020 aagatgtgca cggtgtggag atttatgaag gggatattgt tcaagattgt tattcgagag 37080 aagtaagttt tatcgagttt aaagaaggag ccttttatat aacttttagc aatgtaactg 37140 aattactaag tgaaaatgac gatattattg aaattgttgg aaatattttt gaaaatgaga 37200 tgctattgga ggttatgaga tgacgttcac cttatcagat gaacaatata aaaatctttg 37260 tactaactct aacaagttat tagataaact tcacaaagca ttaaaagatc gtgaagagta 37320 caagaagcaa cgagatgagc ttattgggga tatagcgaag ttacgagatt gtaacaaaga 37380 tctagagaag aaagcaagcg catgggatag gtattgcaag agcgttgaaa aagatttaat 37440 aaacgaattc ggtaacgatg atgaaagagt taaattcgga atggaattaa acaataaaat 37500 ttttatggag gatgacacaa atgaataatc gcgaaaaaat cgaacagtcc gttattagtg 37560 ctagtgcgta taacggtaat gacacagagg ggttgctaaa agagattgag gacgtgtata 37620 agaaagcgca agcgtttgat gaaatacttg agggaatgac aaatgctatt caacattcag 37680 ttaaagaagg tattgaactt gatgaagcag tagggattat ggcaggtcaa gttgtctata 37740 aatatgagga ggaataggaa aatgactaac acattacaag taaaactatt atcaaaaaat 37800 gctagaatgc ccgaacgaaa tcataagacg gatgcaggtt atgacatatt ctcagctgaa 37860 actgtcgtac tcgaaccaca agaaaaagca gtgatcaaaa cagatgtagc tgtgagtata 37920 ccagagggct atgtcggact attaactagt cgtagtggtg taagtagtaa aacgtattta 37980 gtgattgaaa caggcaagat agacgcggga tatcatggca atttagggat taatatcaag 38040 aatgatgaag aacgtgatgg aatacccttt ttatatgatg atatagacgc tgaattagaa 38100 gatggattaa taagcatttt agatataaaa ggtaactatg tacaagatgg aagaggcata 38160 agaagagttt accaaatcaa caaaggcgat aaactagctc aattggttat cgtgcctata 38220 tggacaccgg aactaaagca agtggaggaa ttcgaaagtg tttcagaacg tggagcaaaa 38280 ggcttcggaa gtagcggagt gtaaagacat cttagatcga gttaaggagg ttttggggaa 38340 gtgacgcaat acttagtcac aacattcaaa gattcaacag gacgaccaca tgaacatatt 38400 actgtggcta gagataatca gacgtttaca gttattgagg cagagagtaa agaagaagcg 38460 aaagagaagt acgaggcaca agttaaaaga gatgcagtta ttaaagtggg tcagttgtat 38520 gaaaatataa gggagtgtgg gaaatgacgg atgttaaaat taaaactatt tcaggtggag 38580 tttattttgt aaaaacagct gaaccttttg aaaaatatgt tgaaagaatg acgagtttta 38640 atggttatat ttacgcaagt actataatca agaaaccaac gtatattaaa acagatacga 38700 ttgaatcaat cacacttatt gaggagcatg ggaaatgaat cagctgagaa ttttattaca 38760 tgacggtagt agtttgatat tacatgaaga tgaattattt aacgaaatag tatttgtttt 38820 ggacaatttt agaaatgatg atgactattt aacgatagaa aaagattatg gcagagaact 38880 tgtattgaac aaaggttata tagttgggat caatgttgag gaggcagatg atgattaaca 38940 tacctaaaat gaaattcccg aaaaagtaca ctgaaataat caaaaaatat aaaaataaag 39000 cacctgaaga aaaggctaag attgaagatg attttattaa agaaattaaa gataaagaca 39060 gtgaatttta cagtcctacg atggctaata tgaatgaata tgaattaagg gctatgttaa 39120 gaatgatgcc tagtttaatt gatactggag atgacaatga tgattaaaaa acttaaaaat 39180 atggatgggt tcgacatctt tattgttgga atactgtcat tattcggtat attcgcattg 39240 ctacttgtta tcacattgcc tatctataca gtggctagtt accaacacaa agaattacat 39300 caaggaacta ttacagataa atataacaag agacaagata aagaagacaa gttctatatt 39360 gtattagaca acaaacaagt cattgaaaat tccgacttat tattcaaaaa gaaatttgat 39420 agcgcagata tacaagctag gttaaaagta ggcgataagg tagaagttaa aacaatcggt 39480 tatagaatac actttttaaa tttatatccg gtcttatacg aagtaaagaa ggtagataaa 39540 caatgattaa acaaatacta agactattat tcttactagc aatgtatgag ttaggtaagt 39600 atgtaactga gcaagtgtat attatgatga cggctaatga tgatgtagag gcgccgagtg 39660 attacgtctt tcgagcggag gtgagtgaat aatgagaata tttatttatg atttgatcgt 39720 tttgctgttt gctttcttaa tatccatata tattattgat gatggagtga taataaatgc 39780 attaggaatt tttggtatgt ataaaattat agattccttt tcagaaaata ttataaagag 39840 gtagataaaa atgaacgagc aaataatagg aagcatatat actttagcag gaggtgttgt 39900 gctttattca gttaaagaga tttttaggta ttttacagat tctaacttac aacgtaaaaa 39960 aatcaattta gaacaaatat atccgatata tttagattgt tttaaaaagg ctaaaaagat 40020 gattggagct tatattattc caacagaaca gcatgaattt ttagattttt ttgatattga 40080 agtctttaat aatttagata agcaaagtaa aaaagcgtat gaaaatgtta ttggatttag 40140 acaaatgatt aatttatcaa atagagttaa ggcaatggaa gattttaaga tgagtttcaa 40200 caatgaattt agtacaaatc agattttttt taatccttct tttgttatgg aaacaattgc 40260 tattataaat gaatatcaaa aagatatatc ttatttaaaa aatataatta ataaaatgaa 40320 tgaaaataga gcttataatc atattgatag ttttatcact tcagagtacc gacgaaaaat 40380 aaacgattat aatctttatc ttgataaatt tgaagaacag tttagtcaaa agtttaaaat 40440 aaacagaact tcgataaaag aaagaattat tattaattta aacaagagga gatttaaatg 40500 atgtggatta ctatgactat tgtatttgct atattgctat tagtttgtat cagtattaat 40560 agtgatcgtg caagagagat acaagcactt agatatatga atgattatct acttgatgaa 40620 gtagttaaaa ctaaagggta caacgggtta gaagaataca ggattgaatt gaagcgaatg 40680 aataacgata ttaaaaagta atttatatta tcggaggtat tgcattgaat gataaagatt 40740 gagaaacacg atatcaaaaa gcttgaagaa tacattcagc acatcgataa ctatcgaaga 40800 gagttgaaga tgcgagaata tgaattactt gaaagtcatg aaccagataa tgcgggagct 40860 ggcaaaagta atttgccggg taacccgatt gaacgatgtg caataaagaa gtttagtgat 40920 aacaggtaca atacattaag aaatatagtt aacggtgtag atagattgat aggtgaaagt 40980 gatgaggata cgcttgagtt attaaggttt agatattggg attgtcctat tggttgttat 41040 gaatgggaag atatagcaca ttactttggt acaagtaaga caagtatatt acgtagaagg 41100 aatgcactga tcgataagtt agcaaagtat attggttatg tgtagcggac ttttacccta 41160 tgtaagtccg cattaaaaca gtttattatg ttagtatcag attaatattt aaagttatta 41220 aatgctaata cgacgcatga acaagaggcg catcactatg tgatgtgtct ttttatttat 41280 gaggtatgaa catgttcaaa ctaattgtaa atacattact acacatcaag tatagatgag 41340 tcttgatact acttaagtta tataaggtga aacattatga tgactaaaga cgaacgtata 41400 cgattctata agtctaaaga atggcaaata acaagaaaaa gagtgctaga aagagataat 41460 tatgaatgtc aacaatgtaa gagagacggc aagttaacga catatgacaa aagcaagcgt 41520 aagtcgttgg atgtagatca tatattatcg ctagaacatc atccggagtt tgctcatgac 41580 ttaaacaatt tagaaacact gtgtattaaa tgtcacaaca aaaaagaaaa gagatttata 41640 aaaaaagaaa ataaatggaa agacgaaaaa tggtaaatac ccccgggtca aaaaaatcaa 41700 aagcgatc 41708 19 43576 DNA Staphylococcus bacteriophage 19 tctccataaa aatatgcttg gaaaccttga tttaatgggg ttttaatcta gcaagtgtca 60 aatatgtgtc aagaaaataa ttttctgaca cgttgacctt gctctttttt atgttcatca 120 agtaagtgag agtaggtgtc taaagttata gatatattat aatggcctaa tcttttgcta 180 atatattcaa taggtatacc tttagaaagt aggaaagatg tatgcgtgtg tcttaatgaa 240 taaggtgtta ttgtagtatc atttagtcct atttgactct tagcatggtt aaatgacttt 300 ttaacggcat tatgactcaa tttaaacaac ttattatctg tacgttttgg taattttgat 360 aatttagctt taatatgttg tatatccttt tttggtacct ccacaagtct gtccgcgtta 420 actgtttttg ttccacgaag atgtattgta ccctcttttt cgtttagatc gataggcaac 480 atattaatta catcgctgta tcttgcacca gtgatagcta ggatgaataa aaaaatataa 540 ctcgattcgt ctctagattt aaagtattct atcaattgca agtattgttc tatggtgatg 600 aatttagagt gttcgtcttt tgattttttt gtaccacgaa tatctatttg atagctaggg 660 tctttcttta aatagccctc atatactgca tctctgaagc attgtgataa acaactgttt 720 aatttacgaa ccgtttcatt agtacgacct cgaccgaatt cgttcaaaaa cttttgatac 780 tccgaacgtt tgatgttttt tattaaaaaa tcactcccga aatattcgtt aaataatttt 840 aatgaacgtt gataccaata gaattgttgt gaagcgacat gtttcttatt ttttgaatct 900 aaccaatcat tgtaatattc ttcaaacttt ttattttcat ctaaattgtt tccatcatcc 960 aaatctctaa gcagttgttg agcagcgttg gttgcctcag ctttagtttt gaatcctgac 1020 tttcttttct ttcctgattt gaaagacgga tgttttacgt cgtactgcca agatgctgtt 1080 gctttattct tcctttttgt aattgtaaat gacgccattt tacttttcct cctcaaaatt 1140 ggcaaaaaat aataagggta ggcgagctac ccgaaatttt attgttgaac aactattgct 1200 tcacttcttg cttttcctac ttcttttcta aaactatcat atgattgatt agggtgtgtt 1260 aacgacattc ctggaccacc tccagcatgt tggtttttgt ccggattatt ttccatttct 1320 tcagtggctc ttttagcatt taaatattct tcgtaactag gttcgtttgg gtcgcgtggt 1380 tgtgcttgtt gtccattatt ggtagctgga agattcttct gtacctgttg cttagatgtg 1440 ttattggttt gttgattgtt gttaatgttt gtgttgttct cgttgtttac ttgattattg 1500 ttatcgtttt gattactatt ttcttttttc gcttctgctt tatctttagt ttctttcttt 1560 ttgtctttgt tctctttctt tgtttcggtt ttcttgcttt cctctttctt atcgccgtcg 1620 ttgctaccgc atgcacctaa cactaacgca ctagctaata ataaaactaa taatcttttc 1680 atgttttaca ctcctttatt tgctatttgt tttaataaat ctatgatttc attgttttgt 1740 tctatgattt tgttttcatt tttaagatgt tcgtctaaca tctctattaa gacgaaattt 1800 tgatttatca tttcgtaagt aaacatttga cctgtgttgt taggattaga aaacgaacta 1860 ctgaaacgcg ttgaaaagct atctataaat tgaccaactt tattttttaa taacatatct 1920 ttaccgctct cagacattgt atttagttcg cgcttattta aagttttttc tataattttg 1980 tattttgttt cctgatttct ttcgatttct tctacttcaa aagggatatt gttattaaat 2040 ttttcgataa tatcacgttt ttcagaaact gacatacgat caaatacttg tttttgacct 2100 ttatttaact tccctcgaat ttttccggca gtccaagact ctttaactgt taacttatca 2160 ttaggaactt gattcatctt ttatatgact ccttttctca tatttcttta tatttaaaaa 2220 ctctcaacgg ctcaaatgta atcgaatact cgccatagtg agttccaata ccgtatatct 2280 tcttatattg ttctattgcc tccaatatgt attcttcgct taattgtaga tactcagaca 2340 actcatacaa gttacgtacg ccataattgt aagcttctac aatttcgcgt aacgggactg 2400 ctgagataaa gccgtgtcgt cttgcgtaat tttcgaactt gcgattgttg aatttcgatt 2460 gatctaaaat gttgccatac gtcaacttgt ggtgggcaag ttcttcatat aatacttcta 2520 atttgttcct ttcggataag gaaggtctaa taaaaatttc tccttcttga taccaaccat 2580 cgaatcctcg aggtactctt tgtgtttctt tcacttcaac ttcacatttc ataagcaatt 2640 cttcgtattt tcccatgcgc caaacccctt tggtgtctta tttctttcta tctctaaccc 2700 attgcataaa attttcgatt tcttcccatt cttcgggagt aaattcatct ttatttgcat 2760 gaccggctat agtttcttga tgaatacttc tttcttctgt aattctcgat ttaggtacat 2820 taaagtaatc tgctaattgt tggacttttg atattctagg atatttaagt tctttaagcc 2880 agttagagat tgttgattga cttaccccga ttgcttcaga caattctact tgagtaatgt 2940 tgttctcttt cataagttgt tctaagttct ctgataaaat ttttctagca ctcttatatt 3000 ccataatttt ctcctttagt attacttaat gtaatactaa tttaccataa gtaatatcac 3060 ttttcaatac aaaatattac ttttttgaaa taaatatcac tttaggtgtt gacatattac 3120 tttaagtgat agtatagttg taaatgtcaa cgggaggtga tacgaaatgc cagaaaattt 3180 taaagagttc tctgtaaagg tctggagaac taattcgaat atgacacaac aagatgtcgc 3240 tgataaatta ggcgttacta aacaatctgt aataagatgg gaaaaagatg acgcagaatt 3300 aaaaggctta caattgtatg ctttagccaa attattcaac acagaagttg attatataaa 3360 ggctaaaaaa atttaacatt aatatcactt taagtgataa aggaggaaac tgaaatgcaa 3420 gaattacaaa catttaattt tgaagaatta ccagtaagga aaattgaagt ggaaggagaa 3480 cccttctttt taggtaagga tgttgctgaa attttagggt atgcacgagc agataacgcc 3540 atacgcaatc atgttgatag tgaagatagg ctgatgcacc aaattagtgc gtcaggtcaa 3600 aacagaaata tgatcatcat caacgaatct ggattataca gtttaatctt tgacgcttct 3660 aaacaaagta aaaacgaaaa cattagagaa accgctagga aattcaaacg ctgggtaact 3720 tcggaagttt taccgacgtt aagaaaaact ggtgcttacc aagtacctag tgacccaatg 3780 caagcattga gattaatgtt tgaagctaca gaagaaacaa aacaagaaat taaaaacgtg 3840 aaagatgatg ttattgattt gaaagaaaat caaaaactgg atgcgggaga ctacaatttc 3900 ttaactagaa caatcaatca aagagtagct catatacaaa gactacatgc gataacaaac 3960 caaaaacaac gtagcgaatt attcagggat attaattcag aagtgaaaaa gatgactggt 4020 gcgagttcaa gaacgaacgt aagacaaaaa catttcgacg atgtaattga aatgattgct 4080 aattggttcc cgtcacaagc tactttatac agaatcaagc aaattgaaat gaaattttaa 4140 aacgaaatat aggagaggct gaatatggaa tacatcggat atgcagacgc aaatgcgttt 4200 gtaaaaataa gtggcatttc aaaagatgat ctagagaaaa aagtctactc gaacaaagag 4260 tttcaaaaag aatgcatgta cagatttggt cgaggacaaa agcgttatat aaaaattgac 4320 aaagctattc aatttatcgg taccaattta atgattaatg aatacgaatt ataggaggag 4380 ttatcaaatg agtaaaactt ataaaagcta cctagtagca gtactatgct tcacagtctt 4440 agcgattgta cttatgccgt ttctatactt cactacagcg tggtcaattg caggattcgc 4500 aagtatcgca acattcatat actacaaaga atacttttat gaagaataaa aaaactgcta 4560 cttgcgtcaa caagtaacag tgacaaacat ttatcaaaat atacaactta attaaatcaa 4620 aatatacgga ggtagtcaac tatggctgaa aatattaaaa ctgaacaaca ttattacact 4680 aaagatttct caggatacag aaatgaagaa gataactttg tagcaaatca agaattgaca 4740 gtaacaatca cattgaacga gtacagaaaa cttattgaaa taaaggctgt taaagataaa 4800 gaagaagata cttacagagg taagtatttt gcggaagaaa gaaaaaacga aaaattggaa 4860 aaagaaaata taaaactaaa aaacaaaatt tatgaattac aaaacgaaga agataacgag 4920 gaggacgaag aagacaagga ggacgagaac gatgtattac aaaattggtg agataaaaaa 4980 caaaattata agctttaacg ggtttgaatt taaagtgtct gtgatgaaga gacatgacgg 5040 tatcagtata caaatcaagg atatgaataa tgttccactt aaatcgtttc atgtcataga 5100 tttaagcgaa ctatatattg cgacggatgc aatgcgtgac gttataaacg aatggattga 5160 aaataacaca gatgaacagg acaaactaat taacttagtc atgaaatggt aggaggtatg 5220 aaaagtgaat gatttacaag agagagaatt agaaacattc gaacaagacg accgattcaa 5280 agtaactgat ctagacagtg ctaactgggt ttttaagaaa ctggatgcaa tcacaactaa 5340 agagaatgaa atcaacgatt tagcaaataa agaaattgaa cgcataaacg aatggaaaga 5400 taaagaagta gaaaaattac agagtggcaa agaatattta caaagccttg taattgaata 5460 ttacagaata caaaaagaac aagatagcaa attcaagttg aatacacctt acggaaaagt 5520 gacagccaga aaaggttcaa aagtcattca agttagcaat gagcaagaag tcattaaaca 5580 acttgagcaa cgaggttttg acaactatgt aaaagtaact aaaaaactta gccaatcaga 5640 cattaagaaa gatttcaatg taactgaaaa cggcacattg attgacgcaa acggcgaagt 5700 tttagagggt gctagcattg tggagaaacc aacgtcatac acggtaaagg tgggagaata 5760 gatgactgaa aaaactaatc aagatgtcga tattttaacg caactaggtg taaaagacat 5820 cagcaaacaa aatgcaaaca agttttataa atttgcgata tacggcaagt tcggtactgg 5880 taaaactacg tttttaacaa aagataacaa taccttagta ctagatataa atgaggacgg 5940 aacaacggta acagaagatg gggcagttgt gcagattaag aattataagc attttagtgc 6000 agtgattaaa atgctgccta aaattattga acaactaaga gaaaacggaa aacaaattga 6060 tgttgtagtg attgaaacaa tccaaaagtt acgtgatatc actatggacg acatcatgga 6120 cggtaaatca aagaaaccga catttaatga ttggggcgag tgtgctacac gcattgtaag 6180 tatttatcgt tatatttcta aattacaaga acattatcaa tttcatcttg ctataagcgg 6240 acacgagggc attaacaaag acaaagatga tgagggaagt actatcaatc caacaatcac 6300 gatagaggca caagaccaaa taaaaaaagc agtcatcagt caatctgacg tgttagcaag 6360 aatgacaata gaagaacatg agcaagacgg cgaaaaaact tatcaatatg tacttaacgc 6420 tgaaccatca aatttattcg agacaaagat aagacactca agcaacatca aaattaacaa 6480 caaacgtttc attaatccaa gtattaacga tgttgtacaa gcaattagaa atggtaatta 6540 aaaattaatt aaaaggacgg tataaaaatt atgaaaatca ctggtagaac acaatacatt 6600 caagaaacta atcaagaggc attcatgaaa ggtggggact ttttaggagc tggagaattt 6660 acagtaaaag ttgcaaatgt cgagtttaac gacagagaaa acagatactt cacgattgtt 6720 tttgaaaaca acgaaggtaa acaatacaaa cacaaccaat tcgtcccacc attccaacaa 6780 gattatcaag aaaaacaata tatcgagtta cttagtagat taggaattaa attgaactta 6840 ccagatttaa cttttgacac agatcaatta attaacaaaa tcggaactat tgtacttaaa 6900 aataaattta acgaggaaca aggcaagtat tttgtaagac tctcatatgt aaaagtttgg 6960 aataaagacg atgaagtagt taataaacca gaacctaaaa ctgatgagat gaaacaaaaa 7020 gaacagcaag caaatggtaa acagacacct atgagtcaac aatcaaaccc attcgctaat 7080 gctaatggtc caatagaaat caatgatgat gatttaccgt tctaggacgt ggtttaaatg 7140 caatacatta caagatacca gaaagacaat gacggtactt attccgtcgt tgctactggt 7200 gttgaacttg aacaaagtca cattgattta ctagaaaacg gatatccgct aaaagcagaa 7260 gtagaggttc cggacaataa aaaactatct atagaacaac gcaaaaaaat attcgcaatg 7320 tgtagagata tagaacttca ctggggcgaa ccagtagaat caactagaaa attattacaa 7380 acagaattgg aaattatgaa aggttatgaa gaaatcagtc tgcgtgactg ttcaatgaaa 7440 gttgcgagag agttaataga actgattata tcgtttatgt ttcatcatca aatacctatg 7500 agtgtagaaa cgagtaagtt gttaagcgaa gataaagcgt tattatattg ggctacaatc 7560 aaccgcaact gtgtaatatg cggaaagcct cacgcagacc tggcacatta tgaagcagtc 7620 ggcagaggta tgaacagaaa caagatgaat cactacgaca aacatgtgtt agcactgtgt 7680 agacaacatc ataatgaaca gcacgcaatt ggtgttaagt cgtttgatga taaatatcaa 7740 ttgcatgact cgtggataaa agttgatgag aggctcaata aaatgttgaa aggagagaaa 7800 aatgaataag ttactaatag atgactatcc gatacaagta ttaccgaaat tagctgaatt 7860 aatagggtta aacgaagcaa tagtattgca acaaattcat tattggctaa acaactcaaa 7920 acataaatac gatggcaaaa cttggatttt taattcttat ccagaatggc aaaaacaatt 7980 tccattttgg agcgagagaa ctataaaaag gacatttggg agtttagaaa aacaaaattt 8040 attgcatgta ggtaactaca acaaggctgg atttgaccgt acaaaatggt attcaatcaa 8100 ttatgaaaca ttaaacaaac tagtggcacg accatcggga caaaatggcc cgacgatgag 8160 gacaaattgg cacgatgcaa gaggacaaaa tgacccgacc aataccatag actacacaga 8220 gactaacaaa catagagaga cagacgacgt ctcaaagtca tttaagtata ttagtaccaa 8280 tttagaaatt atacaaaacc ctttaaaagc agaacagtta gaacacgaaa ttaaatcatt 8340 taagcaagat cagttcgaaa tagtaaaagt cgctaccgat tactgcaaag aaaacaacaa 8400 aggtctgaat tacttactaa ctgtattaaa gaactggaat aaagaaggcg tttcagataa 8460 agaaagtgct gaaaacaaat tgaaacctcg taactctaaa aaagaaacta ctgatgatgt 8520 catagcacaa atggaaaaag aattgagtga tgactaatgc cgatgagcaa aacacaagca 8580 ttagaaatta ttaaaaaagt taggtacgta tacaacatcg attttgataa accaaagtta 8640 gaaatgtgga ttgatgtatt aagtcaaaac ggggattatc aaccaactgt aaaagctgta 8700 gatggatata tcaacagtaa caacccgtac ccgcctaacc taccagcaat catgcgtaag 8760 gcacctaaaa aagtatctat tgagccggta gacaacgaaa ccgctacaca ccaatggaaa 8820 atgcagaatg accccgaata tgtcagacaa agaaaaatag cgctagataa cttcatgaat 8880 aagttggcag aatttggggg cgataacgaa tgaattacgg tcaatttgaa attgaaagca 8940 caataatcgc tacgctactt aaacaaccgg acgtactaga aaagataaga gttaaagatt 9000 acatgtttac gaacgaaaag tttaaaacct ttttcaatta tgtaatggac gtcggaaaga 9060 tagatcatca agaaatctat ttaaaagcaa ctaaagataa agagttttta gatgcagata 9120 ctataactaa actttacaac tccgatttca ttggatacgg attctttgaa cgttatcaac 9180 aagaattatt ggaaagttat caaatcaaca aagcgaaaga attggtaact gagttcaaac 9240 aacaacctac gaaccaaaat tttaataact tgattgatga actcaaggat ttaaaaacaa 9300 ttactaacag aaaagaagac ggaaccaaga agtttgttga ggagtttgtc gatgagttat 9360 acagcgatag ccctaagaag caaattaaga cgggttataa gctcatggat tacaaaatag 9420 ggggattgga gccgtcgcaa ttaatcgtca tcgcagcgcg tccctcagtg ggtaagacag 9480 gttttgcatt aaacatgatg ctgaacatag cacaaaatgg atacaaaaca tctttcttta 9540 gtctcgaaac aactggcaca tcagtattga aacgtatgtt atcaacaatt actggtattg 9600 agttaacaaa gataaaagaa atcaggaact taacgccgga tgacttaaca aagttaacga 9660 atgcgatgga taaaatcatg aaattaggca tcgatatttc tgataaaagt aatatcacac 9720 cgcaagatgt gcgagcgcaa gcaatgaggc attcagacag gcaacaagtt atttttatag 9780 attatcttca actgatggat actgatgcga aagttgatag acgtgtagca gtagaaaaga 9840 tatcacgtga cttaaagata atcgctaacg agacaggcgc aatcatcgta ctactttcac 9900 aactgaatcg tggtgtcgag tctagacagg ataaaagacc aatgctatcg gacatgaaag 9960 aatcaggcgg aatagaagca gatgcgagtt tagcgatgct actttaccgt gatgattatt 10020 ataaccgtga cgaagatgac agtatcactg gcaaatctat tgttgaatgt aacatagcca 10080 aaaacaaaga cggcgaaacc ggaataattg aatttgagta ttacaagaag actcagaggt 10140 ttttcacatg aatataatgc aattcaaaag cttattgaaa tcgatgtatg aagagacaaa 10200 gcaaagcgac ccgattgtag caaatgtata tatcgagact ggttgggcgg tcaatagatt 10260 gttggacaat aacgagttat cgcctttcga tgattacgac agagttgaaa agaaaatcat 10320 gaatgaaatc aactggaaga aaacacacat taaggagtgt taaaaaatgc cgaaagaaaa 10380 atattactta taccgagaag atggcacgga agatattaag gtcatcaagt ataaagacaa 10440 cgtaaatgaa gtttattcgc tcacaggagc ccatttcagc gacgaaaaga aaattatgac 10500 tgatagtgac ctaaaacgat ttaaaggcgc tcacgggctt ctatatgagc aagagctagg 10560 attgcaagca acgatatttg atatttagag gtggcacaat gagtaaatac aatgctaaga 10620 aagttgagta caaaggaatt gtatttgata gcaaagtaga gtgcgaatat taccaatatt 10680 tagaaagtaa tatgaatggc actaactatg atcgtatcga aatacaaccg aaatttgaat 10740 tacaacctaa attcgggaaa caaagaccga ttacgtatat agccgatttc tctttgtgga 10800 aggaagggaa actggttgaa gttatagacg ttaaaggtaa ggcgactgaa gttgccaaca 10860 tcaaagcgaa gatattcaga tatcagtata gagatgtgaa tttaacgtgg atatgtaaag 10920 cgcctaaata cacaggtcaa gaatggatgg tatatgagga cttagtgaaa gtcagacgta 10980 aaagaaaaag agaaatgaag tgatctaatg caacaacaag catatataaa cgcaacaatt 11040 gatataagaa tacctacaga agttgaatat cagcattacg atgatgtgga taaagaaaaa 11100 gatacgctgg caaagcgctt agatgacaat ccggacgaat tactaaagta tgacaacata 11160 acaataagac atgcatatat agaggtggaa taaatgaagt tgaacgaagt attcgcaact 11220 aatttaaggg taatcatggc tagagataac gtaagtgtcc aagatttgca caatgaaact 11280 ggcgtatcaa gatcaactat tagtggatat aaaaacggaa aagctgagat ggttaactta 11340 aatgtattag ataaattggc agatgctcta ggtgttaatg taagtgaact atttactaga 11400 aatcacaaca cgcacaaatt agaggattgg attaaaaaag taaatgtata gaggtggaat 11460 aaatgagtat cgtaaagatt aacggtaaac catataaatt taccgaacat gaaaatgaat 11520 tgataaaaaa gaacggttta actccaggaa tggttgcaaa aagagtacga ggtggctggg 11580 cgttgttaga agccttacat gcaccttatg gtatgcgctt agctgagtat aaagaaattg 11640 tgttatccaa aatcatggag cgagagagca aagagcgtga aatggttagg caacgacgta 11700 aagaggctga actacgtaag aagaagccac atttgtttaa tgtgcctcaa aaacattctc 11760 gtgatccgta ctggttcgat gtcacttata accaaatgtt caagaaatgg agtgaagcat 11820 aatgagcata atcagtaaca gaaaagtaga tatgaacaaa acgcaagaca atgttaaaca 11880 accggcgcat tacacatacg gcaacattga aattatagat tttatcgaac aggttacggc 11940 acagtatcca cctcaactag cattcgcaat aggtaatgca atcaaatact tgtctagagc 12000 accgttaaag aatggtcatg aggatttagc aaaggcgaag ttttacgtcc aaagagcttt 12060 tgacttgtgg gagggttaac gatggcaacg caaaaacaag ttgattacgt aatgtcatta 12120 caggaacaat tgggattaga agactgtgaa aaatatacag acgaacaagt taaagctatg 12180 agtcataaag aagttagcaa tgtgattgaa aactataaga caagcatatg ggatgaagag 12240 ctatataacg aatgcatgtc gtttggtctg cctaattgtt aaaaggagtg atgaccatga 12300 acgatagcgc acgcaaagaa tacttaaacc aatttttcag ctctaagaga tatctgtatc 12360 aagacaacga gcgagtggca catatccatg tagtaaatgg cacttattac tttcacggac 12420 attataaaac gatgtttaaa ggcgtgaaaa agacatttga tactgctgaa gagctcgaaa 12480 tatatataaa gcaacatgat ttggaatatg aggaacagaa gcaaccaact ttattttaga 12540 ggagatggaa ataatggcaa agattaaaag aaaaaagaag atgacgctac tcgaactggt 12600 ggaatgggca tggaacaatc ctgaacaagt tgaaagtaaa gtgtttcaat cagatagaat 12660 gggcacgctt ggagaatgta gcgaagtaca tttttcaact gatgggcatg ggttttatac 12720 aaaagtagta acagataaag atatttttac tgtagaaatc acagaggaag tcactgaaga 12780 tactgagttt gattgtctag tagaactaaa cgatattgaa ggttttgaaa tatatgaaaa 12840 tgattcaatc agagagttga tagacggtac ttccagagcg ttttatatac taaacgaaga 12900 taaaactatg acattaattt ggaaagatgg ggagttggta gtatgatgca aacctataaa 12960 gtatgtcttt gtatcaagtt ctttgcatct aaatgtgatt ataaattaaa gaaacattat 13020 ttcgtgaaaa gtacgaatga ggaaaaagcc acgaacatgg tattaaaact gattcgtaaa 13080 aagctcccgt tcgaaactgc aagcatagaa gtcgaaaaag tggaggcaat ataatgatac 13140 aaccaacaag agaagaatta attaatttca tgaaaaaaca tggagctgaa aatgttgact 13200 ctatcactga tgagcaaagt gcaataagac actttagagc tcaatcaaaa gtttttaaag 13260 acgaacgtga tgagtacaag aagcaacgag atgagcttat cgaggatata gctaagttaa 13320 gaaaacgtaa cgaagagctg gagaacatgt ggcgcacagt caaaaatgaa ttgcttggaa 13380 gatacgaaca ttactgtttt aaaattagag aactacaccc tgagagcaaa gcgaacagga 13440 taggagctct ctatatagga ggtaaaagca ctgcagatat tatactgtcg cgaatggaag 13500 aactagacgg aacaaatgag ttctacgaat ttttagggca aatggaggca gacacaaatg 13560 aataaccgtg aacaaataga acaatcagtg atcagtacta gtgcgtataa cggtaatgac 13620 acagaggggt tactaaaaga gattgaggac gtgtataaga aagcgcaagc gtttgatgaa 13680 atacttgagg gaatgacaaa tgctattcaa cattcagtta aagaaggtat tgaacttgat 13740 gaagcagtag gggttatggc aggtcaagtt gtctataaat atgaggagga gcaggaaaat 13800 gagtattagt gtaggagata aagtatataa ccatgaaaca aacgaaagtc tagagattgt 13860 gcaattggtc ggagatatta gagatacaca ttataaactg tctgatgatt cagttattag 13920 cattatagat tttattacta aaccaattta tctaattaag ggggacgagt gagtggaatg 13980 gaaacgatta aaaaatgtgg tgccgcaccc agttatcaaa aataaaaatt taaagtcggt 14040 atacgtaaca aaagataatg tgaaagaggt tcaaaaagaa ttaggtttct ttgaaatttt 14100 taatgaagaa gtgttattaa ctggattttt atcatttcaa aggataccta tttacattat 14160 ttggattaat cctaaatctc ataagacgcc tagatattac tttgctaacg agcatgagat 14220 tgaaagatat tttgaatttt tggaggacga gtaaatgctt gaaatcatcg accaacgtga 14280 tgcattgcta gaagaaaagt atttaaacga cgactggtgg tacgagctag attattggtt 14340 gaataaacgc aagtcagaaa atgaacagat tgatattgat agagtgctta aatttattga 14400 ggaattaaaa cgataggaga taacgaataa atgaataatt taacagtaga tcaattaaaa 14460 gaacttttac aaatacaaaa ggagttcgac gatagaatac cgactagaaa tttaaatgac 14520 acagtagcta gtatgattat tgaatttgcg gagtgggtta acacacttga gttttttaaa 14580 aattggaaga aacaaccagg taagccatta gatacacaat tagatgagat tgctgattac 14640 ttagctttca gtttgcaatt aactctgact attgttgatg aagaagattt ggaagagact 14700 actgaggtta tggttgattt gattgaaaat gaagttactt tacctaaact acattcagtt 14760 tattttgttc atgtaatgca tacactaaca gaacaatttg taaaaggtat tgataatagt 14820 attgtacaag ttttaataat gccttttttg tacgccaata cttactatac aatcgaccaa 14880 ctcattgacg catacaaaaa gaaaatgaaa aggaaccacg aaagacaaga tggaacagca 14940 gacgcaggaa aaggatacgt gtaaagacat cttagatcga gtcaaggagg ttttggggaa 15000 gtgacgcaat acttagtcac aacattcaaa gattcaacag gacaaccaca tgaacatttt 15060 actgctgcta gagataatca gacgtttaca gttgttgagg cggagagtaa agaaggagcg 15120 aaagagaagt acgagaaaca agttaagata aggagagatg gagatgccaa agaaaacggt 15180 aacgattgat gtagatgaaa acttattagt agtagctagt aatgaaatat cagaactatt 15240 atatgaatat gacagtgagt taatgtcagc tgatgaagat ggcgataata gagatatcga 15300 aaaaaaaaga gacgcattaa aacaagctat acaaattatc gataaattaa catgtcgagg 15360 aggcagacga tgattaacat acctaaaatg aaattcccga aaaagtacac tgaaataatc 15420 aagaaatata aaaataaaac acctgaagaa aaagctaaga ttgaagatga tttcattaaa 15480 gaaattaatg ataaagacag tgaattttac agtcctatga tggctaatat gaatgaacat 15540 gaattaaggg ctatgttaag aatgatgcct agtttaattg atactggaga tggcaatgat 15600 gattaaaaaa cttaaaaata tggattggtt cgatatcttt attgctggaa tactgcgatt 15660 attcggcgta atcgcactga tgcttgttgt catatcgcct atctatacag tggctagtta 15720 ccaaaacaaa gaagtatatc aagggacaat tacagataaa tataacaaga gacaagataa 15780 agaagacaag ttctatattg tgttagacaa caagcaagtc atcgaaaact ctgacttact 15840 attcaaaaag aaatttgata gcgcagacat acaagctagg ttaaaagtag gcgacaaagt 15900 agaagttaaa acgattggtt atagaataca ctttttaaat ttatatccgg tcttatacga 15960 agtaaagaag gtagataaat aatgattaaa caaatattaa gactattatt cttactagcg 16020 atgtatgagc taggtaagta tgtaactgag aaagtatata ttatgacgac ggctaatgat 16080 gatgtagagg cgccgagtga cttcgcaaag ttgagcgatc agtctgattt gatgagggcg 16140 gaggtgtcag agtagatgta tagcaaagag tcaattgtta atatgatagg cacacataaa 16200 atgaagtgta atgtattagc tgatgtaata ccggaatatg atagcaattc aattgcacag 16260 tatggcatac aagcaacgtt gccgaaacca caaggggaaa actcaagtaa agttgaagat 16320 gttgttgtga ggcttgagag agcaaataaa aggtatgctc agatgttaaa agaggttgag 16380 tttataaatc aatcgcaaca gagattggga cacgttgact tttgcttctt agagttattg 16440 aagaaaggtt ataacaggga tgcgattatc aagaagatgc ctaactctaa attaaataga 16500 aacaacttct tagcgcgccg tgatgagtta gcagaaaaga tttatctact acagtgacga 16560 aaatgacaaa aatgacagaa atgacgaaaa tgacactatt tttaaactgt gaattaattt 16620 tatataattg atttgtaaga attatcttaa gacgtggggt aatagccaca ttagatgttc 16680 tcatcgatgt gattgagaag tgacaaacat ataaaagatg atatgttacg ctattaatca 16740 cctactacct gcctatatgg tgggtagttt aattcttgca ttttgagtca taactatttt 16800 cctcctttca catttattga acgtagctcc tgcacaagat gtaggggcat tttttatatt 16860 taaataacta gagtaattaa cgtaaaggcg tgtgatacag tgaaaacaat tgattaaatt 16920 aacaccgaag caagaaaagt ttgtgctagg actcatagag ggcaagagcc aacggaaagc 16980 atatattgac gcagggtatt cgactaaagg taagagtggg gaatatctag ataaagaagc 17040 gagtacactt tttaaaaatc ggaaggtttc cggaaggtac gaaaaattgc gtcaagaagt 17100 agctgaacaa tcaaaatgga cacgccaaaa ggcctttgaa gaatatgagt ggctaaagaa 17160 tgtagctaag aatgacattg aaatagaggg agtgaagaaa gcgacagctg atgcattcct 17220 cgctagttta gatggtatga atagaatgac gttaggtaac gaagttttag ctaaaaagaa 17280 aatagaaact gaaattaaga tgcttgagaa gaagattgaa caaatagata aaggtgacag 17340 tggaacagaa gataaaatca aacaacttca cgacgcaata acggaagtga tcgtcaatga 17400 ataaacttaa atctttatat acggacaaac aaattgaaat attgaagcaa acgcaaaaac 17460 aagattggtt tatgttaatt aatcacggag caaagcgtac aggtaaaaca atattaaaca 17520 atgacttatt tttacgtgag ttaatgcgtg tgcgaaagat agcagacgaa gaaggaattg 17580 agacacctca atatatactt gctggtgcaa cattaggtac gattcaaaaa aacgtactaa 17640 tagagttaac taacaaatat ggcattgagt ttaattttga taaatataat tcattcatgt 17700 tatttggcgt tcaagtggtt cagacaggtc acagtaaagt aagtggtata ggagctatac 17760 gtggtatgac atcgtttggt gcatatatca atgaagcgtc gttagcgcat gaagaggtgt 17820 ttgacgagat taagtcacgt tgtagtggaa ctggtgcaag aatattggta gataccaacc 17880 ctgaccatcc cgagcattgg ttgttgaaag attatattga aaatacagat cctaaagcag 17940 gtatactgag tcaccaattt aagctcgatg acaataactt tcttaatgat agatataaag 18000 agtctattaa ggcttcaaca ccatcaggta tgttctatga acgtaatatc aacggtatgt 18060 gggtgtctgg tgacggtgta gtatatgccg actttgattt gaatgagaat acgattaaag 18120 cagatgaact ggacgacata cctatcaaag aatactttgc tggtgtcgac tggggttacg 18180 agcactatgg atctattgtg ttaataggac gaggtataga tggtaacttt tattttattg 18240 aggagcacgc acaccaattt aagtttattg atgattgggt ggttattgca aaagatattg 18300 taagtagata tggcaatatt aatttttact gcgatactgc acgacctgaa tacatcactg 18360 aatttagaag acatagatta cgtgcaatta acgctgataa aagtaaacta tcgggtgtgg 18420 aggaagttgc taagttgttc aaacaaaaca agttacttgt tctttatgat aatatggata 18480 ggtttaagca agaggtattt aaatatgttt ggcaccctac aaacggagag cctataaaag 18540 aatttgatga cgtgttggac tcgttaagat atgccatata cacacatact aaacctgaac 18600 gattaaggag ggggaaatga cattgtataa gttaatagat gatattgaag cacaaggaat 18660 attgcctaag catattgagg ctctaataga gtcacataaa gacgatagag agagaatggt 18720 taatctctat aatagataca agacacatat tgactatgta ccaatattca aacgtcgacc 18780 aattgaagaa aaagaagatt ttgaaactgg tggaaatgta aggcgattag acgtgtctgt 18840 taataacaaa cttaacaact cttttgacag cgaaattgtt gatacacgtg ttggttattt 18900 acatggtgtt cctgttactt atgatttaga tgaaaacgca gaaaaaaacg aaaagttgaa 18960 aaagtttata accaactttg ccattagaaa tagtgttgat gatgaggatt ctgaaatagg 19020 taaaatggca gcaatttgcg gatatggtgc taggttagca tatattgata cgaatggtga 19080 tattaggatt aagaatatag atccctataa tgttattttt gttggcgaca atattttaga 19140 acctacatac tcattgcgct acttttatga aaaagatgat gataatggca ctgattatgt 19200 gtacgcagag ttttacgata atgcttatta ttatgtattt cgaggagaag gtattgacgc 19260 tttgcaagaa gttggacgat atgaacattt atttgattac aatccattgt ttggtgtacc 19320 taacaacaaa gagatgatag gagatgctga aaaggttatt cacttaattg acgcatatga 19380 tttaacaatg agcgatgcat caagtgagat tagtcagaca cgtttagcat accttgtgtt 19440 acgcggtatg ggtatgagtg aagaaatgat tcaagaaaca caaaagagtg gcgcatttga 19500 gttgttcgac aaagatatgg acgttaaata cttaacaaaa gatgtaaatg acacaatgat 19560 tgagaaccat ttagatcgaa tcgaaaagaa tatcatgcgt tttgcaaagt cagtaaactt 19620 taattctgac gagtttaacg gaaatgtacc tatcattgga atgaaactta aacttatggc 19680 tttagagaac aagtgtatga cgtttgagcg taagatgaca gctatgttga ggtatcaatt 19740 caaagttatt ttatctgcat taaagcgtaa agggtacaac ttggatgatg atagttattt 19800 aaacctgata tttaagttca ctcgtaacat tccagttaat aagttagaag aatcacaagt 19860 gctaattaac ctgaagggac aagtttcaga acgaacaagg ttaggacaat cacaactagt 19920 tgatgatgtt gattacgaat tagacgaaat ggaaaaagaa agtcttgaat ttaatgacaa 19980 attacctgac atagatgaag gtgacgcaaa tgacaaatcc caaaataacc aatcagaatg 20040 atattgatga gtatatcgag ggtttaatct ctaaagcaga aaaaccaata gaacaactat 20100 ttgctaatcg acttaaagag ataaaacaaa tcatcgcaga tatgtttgag aaatatcaaa 20160 atgatgatgt gtatgttaca tggactgaat tcaataaata caacaggctc aataaggagt 20220 taactcgtat aggtacaatg ttgacttatg actataggca agtagctaag atgattcaga 20280 agtcacaaga agatgcttat atagaaaaat tccttatgag cctttattta tatgaaatgg 20340 cgagtcaaac atctatgcag tttgatgttc cgagtaaaga ggtaatcaaa tcagctattg 20400 aacaacctat tgagttcatt cgtttaatgc caacactaca aaaacatcgt gatgaagtat 20460 tgaaaaagat acgtatgcac attacacaag gtattatgag tggagagggt tactctaaga 20520 tagctaaagc aatacgtgat gatgtcggca tgtctaaagc tcaatcattg cgtgtggctc 20580 gtacagaagc aggcagagca atgtcacaag ctggacttga tagcgcaatg gttgctaaag 20640 ataacggttt gaatatgaag aaacgttggc atgctactaa agatacacga acacgtgata 20700 ctcatcgtca tttagatggg gaatcagtgg aaatagatca gaattttaaa tcaagtgggt 20760 gtgttgggca ggcgcccaag ctatttattg gtgtaaacag tgcgaaagag aatattaatt 20820 gtcgttgcaa attactttat tatattgatg aaaatgaatt gccaactgta atgagagcac 20880 gtaaagacga tggtaaaaat gaagttatcc cattcatgac ttatcgtgag tgggagaaat 20940 ataagcgaaa aggtggtaat tgatatggat tttaaaataa aagtaaatgt tgatactggc 21000 gaagctatag aaaagttaga acgcattaaa tccttgtacg aagagataat agagttacaa 21060 aacgaaaaag ttgttgtaaa cgtaacagtt aaaaatgaag ctgatttaga tatggttaaa 21120 acatctatta gcgaagaaaa tgctaaaaat aatgatttca cactttttta gttgtctctt 21180 tgctactcga ccttagcatg tcgttaaact gctttttatt atgcactttt cggactgtta 21240 gggtacgcga agggcaaaaa ggagttttga tatatgaata tcgaagaagt taagtctttt 21300 tttgaagaac acaaagacga taaagaagta aaagattatc taaagggact taagacggtg 21360 tctgttgatg acgttaaagg ctttttagat acagaagaag gtaaacgatt cattcaacct 21420 gaattagatc gttatcattc gaaaggatta gaatcatgga aagagaaaaa tcttgaggat 21480 ctaatcgaac aagaagtacg gaagcgtaat cctgagcaat cagaagaaca aaaacgtatt 21540 agtgctcttg aacaagagtt agaaaaacgc gacgcagagg caaaacgtga gaagttaaga 21600 agtaacgcgc taggtaaagc gcaggaacta aatttaccaa catccttagt tgatagattt 21660 ttaggcgatt ctgatgaaga tactgagcaa aacttaaaag ctttaaaaga aacctttgac 21720 aagtatgttc aaaaaggcgt tgagtctaaa tttaaatcga gtggaagaga tgttaaagaa 21780 tcacgaaatc aagatttaga cccttcaaat gtaaagtcca ttgaagaaat ggcgaaagaa 21840 atcaatatta gaaaataaag tgaggtaata aaatatggca actccaacat acacgccagg 21900 caatgttatt ttatcggatt ttaaaaacgg cgttattcca gcagaacaag gtactttaat 21960 catgaaagac attatggcta attcagcaat tatgaaatta gctaaaaatg agccaatgac 22020 agcacaaaag aaaaaattta cttacttagc aaaaggtgta ggcgcctact gggtatcaga 22080 aacggaacgt attcaaactt ctaagcctga atatgcgcaa gcagaaatgg aagctaagaa 22140 aattggtgta attattccgt tatcaaaaga gtttcttaaa tggactgcaa aagatttctt 22200 taatgaggtt aaacctctaa ttgcagaggc attttacaaa gcgtttgacc aagctgttat 22260 ctttggtact aaatcacctt acaacacttc aactagtggt aaaccgcttg ttgaaggcgc 22320 agaagagaaa ggtaacgttg ttacagatac taataattta tacgtagacc tttcggcatt 22380 aatggctact attgaagatg aagagttaga tccaaacgga gtattaacta cacgttcatt 22440 cagaagtaaa atgcgtaatg ctttagatgc taatgacaga ccattatttg atgctaacgg 22500 gaacgagatt atgggattac cactatctta tactggagcg gatgtatacg acaaaaagaa 22560 atcgttagca ctaatgggtg attgggatta cgcacgttac ggtatcttac aaggtattga 22620 gtatgcaatt tctgaagatg ccacgttaac gacgttacaa gcatcagatg cttctggcca 22680 accagtatca ttatttgaac gtgatatgtt cgctttacgt gcgacgatgc atattgcata 22740 catgaacgtt aaaccagaag cgttcgcaac gcttaaacca actgaatagg aggagatatg 22800 atggctaatc ctgcagaaga gattaaggta aaaaaagaca atatgactat tactgttaca 22860 aagaaggcat ttgactctta ttacagtctt gtcggttaca aagaggttaa atcacgtcgt 22920 actacgtctg ataagagcga gtgataaaaa tgactcttta tgaagatgtt aaacttttac 22980 tcaagaaaaa tggagtggaa gttaaaagtg atgaagaaga aatatttaag atggaagttg 23040 acggaatact agaagatgtt agggatataa caaacaatga ttttatgaaa gatggtcaag 23100 tcatttatcc ttactcaatc aaaaagtatg tcgcagatgt cctagagtat tatcaacgac 23160 ctgaagttaa aaagaattta aagtcaagaa gtatggggac agtgtcgtac acttataacg 23220 atggtgtccc tgattacatt agtggagtat taaacaggta taaacgagca aagtttcatc 23280 cgtttaaacc aataaggtag aggtgttgtt tgtgtttaac ccatacgacg aattccctca 23340 cactatttct attggaagta tcaaaaaagt aggagagtat ccaattatac aagagcgctt 23400 tgtaagcgat aaaacaatta aaggatttat ggatacgcct actacatctg aacaactaaa 23460 atttcatcaa atgtcacaag aatatgacag aaacctatat gtaccttatg acttgccaat 23520 atctaaaaac aatttatttg agtatgaggg tagaatcttt agtattgaag gtgattctgt 23580 agatcagggc ggacaacatg aaattaagtt actacgactt aagcaggtgc catatggcaa 23640 aagttaagta cggtgctgat agcatggttg ttgaattgga taagttcgat aagaaaatag 23700 aagagtgggt taaaaaaggt attgctaaaa caacgacgaa gatttacaac actgctgtag 23760 cattagctcc tgttgactta ggttttttag aagaaagtat tgactttaaa tatttcgatg 23820 gtgggttatc cagtgttata agtgtcggcg cagattatgc aatatacgtt gaatacggta 23880 ctggtatata tgctactggt cctggtggta gtcgtgctac aaagattccg tggagtttta 23940 aaggtgatga cggcgaatgg tacaccacat atggtcaagc gccacagcca ttttggaacc 24000 ctgcaattga cgcaggacgc aagacattcg agcagtattt ttcatagagg tggttaaata 24060 tgtgggtatc agttgagcct gaacttacaa atcaaatata taaaagatta atctcagacc 24120 ctaacattaa caaactagtt gatgataggg tttttgacgt tgttcaagat gacgctgttt 24180 acccatatat tgttgtgggt gaatcaaacg tcactaacaa cgaatctagc gcaacaatga 24240 gagaaacagt cggtattgtc atacatgtgt attcacagtt cgctacacaa tacgaggcta 24300 agctcatttt aagcgcgata ggttatgtgc ttaacagacc tatagaaata gataattacg 24360 agtttcaatt tagccgtatc gatagtcaag cagtattccc tgatatagac aggtttacta 24420 agcatggcac gatacggctt ttatttaagt acagacataa aaagaaaaac gaaggagtgt 24480 attaaatggc gcaaaaaaac tatttagcag ttgtacgtcc agctgaaact gacttagatc 24540 cagtagaatc tttattatta gctgacttac aagaaggtgg acatacgatt gaaaatgatt 24600 tagctgaaat agtacgaggc ggtaaaacgg actattctcc caatgcaatg tcagaatcat 24660 ttaaattaac aattggtaat gtgcctggag ataaaggaat tgaagcagtg aaacacgctg 24720 tacaaacagg tggacagttg cgtatatggc tttatgagcg taataaacgt gcagacggta 24780 aacatcacgg aatgtttggt tatgttgttc cagaatcatt tgaaatgtca tttgatgatg 24840 aaagtgacaa aatcgaacta tcattaaaag ttaaatggaa tacagcagaa ggtgctgaag 24900 ataacttgcc gaaagagtgg tttgaagctg caggtgcgcc tacagttgaa tacgaaaaat 24960 tcggcgaaaa agtcggaaca ttcgagaatc aaaagaaagc tagtgttgta tctgattcac 25020 acacggaaga ccattctatg taaactaata gatcaagggg gcgtaagctc cctatttttt 25080 tataaaaaaa ttgaaaagag gtatatattt tgactgaatt taatccaatt acaacattaa 25140 aaattaatga cggagaaaaa gattacgaag tagaagcaaa agtaacattt gcatttgacc 25200 gaaaagctga aaaattctca gaagatagcg aagatgggag aaaaggagca atgccaggat 25260 tcaatgttat ctttaacggt ttgctagaat ctagaaacaa agcgatttta caattttggg 25320 aatgtgctac tgcttattta aaaaacccac caactcgaga acaattagaa aaagcaattg 25380 atgatttcat cactgaaaac gaggatactt tgccgttatt acaaggggct ttggacaaac 25440 ttaacaatag tggttttttc aagagggaga gtcgctcgta ctggatgaca ttgaacaaag 25500 caccgaatat ggccaaaagc gaggacaaag aaatgacgaa agcaggcata gaaatgatga 25560 aagagaatta caaggaaatc atgggcgcag aaccttacac gattactcaa aaataaggca 25620 actgacagct agatatttag gatatatccc tgaacatgaa ttgttagcac taacacctgc 25680 tgaatggcgt gattggctta ttggtggtca ggataggtac ctagatcaaa gacaattatt 25740 aattgaacaa gcgcaagcta acggcttagt acaagcttct aagaggctaa ctagtatgat 25800 tcgtgacatt gagaaacaac gttacgaaat aagagaacct ggtagctatg ctcgtgtaca 25860 aaaagctaga ttagaagaag aaaaaagaag acgtgaactc ttcaaagaag gtacaagaaa 25920 attccttgaa tcgaaaggag gttagccttt ggatactcat tttatggcaa agattatggc 25980 caatattaga gatttccaaa gcaacgtaag gaaagctcaa cgattagcaa agacgtctgt 26040 accaaacgaa attgaaacag atgtaaaagc agatatttca agattccaaa gagctttaca 26100 acgcgctaaa tcaatggctc aacgatggcg agagcattct gttaaattat tcatgaaaac 26160 agatgagtat aaagcgaatt tagaacgcgc taaagctcaa gtagagcgat ttaaacaaca 26220 taaagtagat ttgaaactaa gtaacactga attaatggcc aaatataatg caactaaagc 26280 tactgtcgaa gcttggagaa aacatgttgt taagttggat ttagatgcaa accccgctaa 26340 aatggcggtt aaagggttta aagaagattt aatagatctt agcaggcata gttttgatat 26400 tgattccagc agatggaaat taggaaataa attcacaaaa gaattcaatg aagtcgaagg 26460 agcagttaaa cgttctttcg gaagaattgg tcagattatg agaaaagaag taaatggaac 26520 aagtgatatt tggggtaaac ttaacaactc attgaaagat tacggcgaga aaatggacgc 26580 cttagctact aaaatccgaa ctttcggtac tatcttcgcg caacaggtca aaggcttaat 26640 gattgctagt atacaagcat tgataccagt gattgccgga ttagtacctg caataatggc 26700 agtacttaat gcggttggtg tattaggtgg tggcgtttta ggtttagttg gcgcattctc 26760 tgtcgcaggt cttggagttg ttggctttgg tgcaatggct attagcgctc ttaaaatggt 26820 tgaagatgga acattggcag taacaaaaga agttcaaaac tttagagatg cgagcgatca 26880 gttaaaaact acatggcgtg atattgttaa agagaatcaa gcaagtatct ttaatgcgat 26940 gtcagcaggt atcagaggcg ttacaagtgc gatgtctcaa ttaaaaccat tcttatccga 27000 agtatctatg ctagttgaag caaacgcacg cgagtttgag aattgggtta aacattccga 27060 aacagctaag aaagcgtttg aagcattgaa tagcataggt ggcgcaatct tcggagattt 27120 attgaacgct gcaggacgat ttggcgacgg attagttaac attttcactc aattaatgcc 27180 gttgttcaaa tttgtgtctc aaggactaca gaacatgtct atagctttcc aaaattgggc 27240 taatagtgta gctggtcaga atgctattaa agcgtttatt gactacacta ccactaactt 27300 acctaagatt ggtcagatat ttggtaatgt gttcgctggt attggtaatt taatgattgc 27360 ttttgcacaa aacagttcca acatttttga ttggttggtt aaattaactt ctcaatttag 27420 agcatggtca gaacaagtag gacaatcaca agggtttaaa gactttatca gttatgttca 27480 agagaatggt cctactatta tgcagttaat cggtaatatc gtaaaagcat tagttgcttt 27540 tggtactgca atggctccta tagctagtaa attgttagac tttatcacta atctagctgg 27600 atttatcgct aaactattcg aaacacaccc agctatagca caagttgctg gcgttatggg 27660 tattttaggc ggtgtatttt gggctttaat ggctccgatt gttgctataa gtagtgtact 27720 tacaaatgtg tttggtttga gcttattcag cgtcactgaa aagattttag acttcgttag 27780 aacatcaagt ttagttactg gagctacgga agcattaata ggtgcattcg gttcgatttc 27840 agcacctatt ttagcagttg ttgcagtaat tggtgcattc attggtgtcc tcgtttattt 27900 atggaaaaca aacgagaact ttagaaatac tattactgaa gcgtggaacg gtgttaaaac 27960 ggcagtttct ggtgcgattc aaggtgtagt cggctggtta actgaattgt ggggcaaaat 28020 ccaatctacc ttacaaccga taatgcctat attgcaagta ttaggacaaa tattcatgca 28080 agttttaggt gttttggtaa taggcatcat tacaaacgtt atgaatatca tacaaggttt 28140 gtggacttta attacaattg cgttccaagc cataggaaca gtgatatccg tagcagtcca 28200 aatcatagta ggtttgttca ctgctttaat tcagttgctt actggcgact tctcaggtgc 28260 ttgggagact attaaaacta cggttaccaa tgtgcttgat acgatttggc aatacatgca 28320 atcagtttgg gagtcaatta tcggcttttt aactggcgta atgaatcgaa cactttctat 28380 gtttggtaca agttggtcac agatatggag tacaatcact aattttgtta gcagtatttg 28440 gaacactgtt acaagttggt tcagtcgagt ggcttcgagt gtagctgaaa aaatggggca 28500 agcactaaac tttattatca caaaaggttc tgaatgggtt tctaacattt ggaatacagt 28560 tacaagtttc gcgagtaaag tagctgatgg gtttaaaaga gttgtctcaa atgtaggtga 28620 cggtatgagt gatgcacttg gtaagattaa aagtttcttc agtgatttct taaatgccgg 28680 agcggaatta atcggcaaag tagctgaggg tgtagccaat gctgcgcaca aagtagtcag 28740 cgcggtaggc gatgcgattt catcagcttg ggactctgta acttcattcg taagtggaca 28800 cggtggaggt agtagcttag gtaaaggttt agcggtatca caagcaaaag taattgctac 28860 agactttggc agtgccttta ataaagagct atcctctact ttgacagata gtatagtaaa 28920 tcctgtaagt acttctatag acagacacat gactagcgat gttcaacata gcttaaaaga 28980 aaataataga cctattgtga atgtaacgat tagaaatgag ggcgaccttg atttaattaa 29040 atcacgcatt gatgacatga acgctataga cggaagtttc aacttattat aagggaggtt 29100 tgttagttga tagcgcacga tatagaagta ataaggaatg gttcacagta tcgcgtcagt 29160 gacaatcctt tcacttataa tcacttggaa gtagttgaat ataacgttac aggcgcagga 29220 tatcatcgta actattctga tatagagggt attgatggta gatttcataa ttacgctaaa 29280 gaagaactta aaaaagtaga gcttaagata aggtataaag tacctaaaat tgcttatgct 29340 tcacatttaa agtcagacgt ccaagcacta tttgctggac gtttttattt aagggaatta 29400 gctacaccag acaattcaat taagtatgag catatattag atataccaaa agacaaacaa 29460 gcatttgagc ttgattatgt tgatggacga caactttttg taggactagt aagtgaagtt 29520 tcttttgaca caacacaaac atcaggggaa ttttctttgt cgtttgaaac aaccgaacta 29580 ccatactttg aaagtgtcgg ttatagtact gatcttgaaa gtaataacga ccctgaaaaa 29640 tggtcggtac ctgatagatt gcctacaaac gaaggtgata agaggcgtca aatgacattt 29700 tacaacacta actcaggaga agtttattat aacggtgatg ttcctttaac acagtttaat 29760 cagtttaatg ttgttgaaat agagttagct gaagatgtta aagctaatga taaggatgga 29820 ttcactttct atacagataa aggaaatatc tcagttatta aggaagttga tttaaaagcc 29880 ggagataaaa taatcttcga cggtaaacat acctatagag gttatttaaa tatagattct 29940 tttaataaaa ctttagaaca accggtttta tatccaggct ggaatcgatt caagtctaat 30000 aaagtaatga aacaaattac atttagacac aaattatatt ttagataagg agtagcctat 30060 gccaatttta ttaaaaagtc tacagggtgt agggcacgct attaatgtta gtacaaaggt 30120 aagtaaaaag ctaaatgaag atagttcttt ggatctaact attatcgaga acgcgagtac 30180 gtttgacgca ataggtgcta taactaaaat gtggacgatc actcatgttg aaggtgaaga 30240 tgatttcaac gaatatgtaa ttgtcatact tgataagtct actattggcg aaaaaataag 30300 gcttgatatc aaagctaggc aaaaagaact tgatgacctt aacaattcta ggatttacca 30360 agagtataac gaaagtttta caggcgttga gttcttcaat actgtcttta aaggaacggg 30420 ttataagtat gtattacatc caaaagtaga tgcatctaaa ttcgagggat taggcaaagg 30480 agatacacga ttagaaatct ttaaaaaagg acttgagcgt tatcatctcg aatatgaata 30540 cgatgcaaag actaaaacgt ttcatttgta tgatgaatta tctaagtttg ccaattatta 30600 cattaaagct ggtgtgaatg ctgataacgt caaaatacaa gaagatgcat ctaaatgtta 30660 tacctttatt aaaggttatg gtgattttga tggacaacag acttttgcag aagcgggact 30720 acaaattgaa ttcactcatc cattagcaca attgataggt aaaagagaag cgccaccgct 30780 tgttgatgga cgtattaaaa aagaagatag tttaaaaaaa gcaatggagt tattgataaa 30840 gaaaagtgtc actgcttcta tttccttaga ctttgtagcg ttacgtgaac atttcccaga 30900 agctaaccct aaaataggtg atgttgttag agtggtggat tctgccatag gatataacga 30960 cttagtgaga atagtcgaaa tcactacaca tagagatgcg tacaataata tcactaagca 31020 agatgtagta ttaggagact ttacaaggcg taatcgttat aacaaagcag ttcatgatgc 31080 tgcaaattat gttaaaagcg taaaatctac aaaatccgac ccatctaaag aactaaaagc 31140 attaaacgca aaagttaacg caagtttatc tataaataat gaattggtta agcagaatga 31200 aaaaataaac gctaaagtcg ataagatgaa tactaaaaca gttacaactg ctaatggtac 31260 gatcatgtac gactttacta gtcaatcaag tataagaaac atcaaatcaa ttggaacgat 31320 tggcgactct gtagctagag ggtcgcacgc aaaaactaat ttcacagaaa tgttaggcaa 31380 gaaattgaaa gctaaaacga ctaatcttgc aagaggtggc gcaacaatgg caacagttcc 31440 aataggtaaa gaagcggtag aaaacagcat ttatagacaa gcagagcaaa taagaggaga 31500 cctaatcata ttacaaggca ctgatgatga ctggttacac ggttattggg caggcgtacc 31560 gataggcact gataaaacgg atacaaaaac gttttacggt gccttttgtt ctgcaattga 31620 agttattaga aagaataatc cagattcaaa aatactagtg atgacagcta caagacaatg 31680 ccctatgagt ggtacaacaa tacgccgtaa agacacggac aaaaacaaac tagggttaac 31740 acttgaggac tatgtaaacg ctcaaatatt agcttgtagt gagttagatg taccagtgtt 31800 tgacgcatat cacacagatt actttaagcc atacaatcca gcttttagga aagcgagcat 31860 ggaggacggc ttacacccta acgaaaaagg tcacgaggtt attatgtacg agttaatcaa 31920 ggattattac agtttttacg actaaaggag gcaaccaatg gcttacggat taattacaag 31980 tttacattca atgacaggtc ggaaaatagt tgctcaacat gagtataact atcgcttgtt 32040 agatgaaggt atgagcaaac ttgagaaaat gtttatatac catcaaaaag aagaaatata 32100 cgcacactca gcgaaacaaa ttaaatactt gaatgacagt gttgaagatt atttaacgta 32160 tttaaatagc cgttttagca atatgattct aggccataac ggcgacggta tcaatgaagt 32220 aaaagacgcg cgtattgata atacaggtta tggtcataag acattgcaag atcgtttgta 32280 tcatgattat tcaacactag atgctttcac taaaaaggtt gagaaagctg tagatgaaca 32340 ctataaagaa tatcgagcga cagaataccg attcgaacca aaagagcaag aaccggaatt 32400 tatcactgat ttatcgccat atacaaatgc agtaatgcaa tcattttggg tagaccctag 32460 aacgaaaatt atttatatga cgcaagctcg tccaggtaat cattacatgt tatctagatt 32520 gaagcccaac ggacaattta ttgatagatt gcttgttaaa aacggcggtc acggtacaca 32580 caatgcgtat agatacattg atggagaatt atggatttat tcagctgtat tggacagtaa 32640 caaaaacaac aagtttgtac gtttccaata tagaactgga gaaataactt atggtaatga 32700 aatgcaagat gtcatgccga atatatttaa cgacagatat acgtcagcga tttataatcc 32760 tatagaaaat ttaatgattt tcagacgtga atataaagct tctgaaagac aagctaagaa 32820 ttcattgaat ttcattgaag taagaagtgc tgacgatatt gataaaggta tagacaaagt 32880 attgtatcaa atggatatac ctatggaata cacttcagat acacaaccta tgcaaggtat 32940 cacttatgat gcaggtatct tatattggta tacaggtgat tcgaatacag ccaaccctaa 33000 ctacttacaa ggtttcgata taaaaacaaa agaattgtta tttaaacgac gtatcgatat 33060 tggcggtgtg aataataact ttaaaggaga cttccaagaa gctgagggtc tagatatgta 33120 ttacgatcta gaaacaggac gtaaagcact tttaataggg gtaactattg gacctggtaa 33180 taacagacat cactcaattt attctatcgg ccaaagaggt gttaaccaat tcttaaaaaa 33240 cattgcacct caagtatcga tgactgattc aggtggacgt gttaaaccgt taccaataca 33300 gaacccagca tatctaagtg atattacgga agttggtcat tactatatct atacgcaaga 33360 cacacaaaat gcattagatt tcccgttacc gaaagcgttt agagatgcag ggtggttctt 33420 ggatgtactg cctggacact ataatggtgc tctaagacaa gtacttacca gaaacagcac 33480 aggtagaaat atgcttaaat tcgaacgtgt cattgacatt ttcaataaga aaaacaacgg 33540 agcatggaat ttctgtccgc aaaacgccgg ttattgggaa catatcccta agagtattac 33600 aaaattatca gatttaaaaa tcgttggttt agatttctat atcactactg aagaatcaaa 33660 acgatttact gattttccta aagactttaa aggtattgca ggttggatat tagaagtaaa 33720 atcgaataca ccaggtaaca caacacaagt attaagacgt aataacttcc cgtctgcaca 33780 tcaattttta gttagaaact ttggtactgg tggcgttggt aaatggagtt tattcgaagg 33840 aaaggtggtt gaataatgat agtagataat ttttcgaaag acgataactt aatcgagtta 33900 caaacaacat cacaatataa tccaattatt gacacaaaca tcagtttcta tgaatcagat 33960 agaggaactg gtgttttaaa ttttgcagta actaagaata acagaccgtt atctataagt 34020 tctgaacatg ttaaaacatc tatcgtgtta aaaaccgatg attataacgt agatagaggc 34080 gcttatattt cagacgaatt aacgatagta gacgcaatta atgggcgttt gcagtatgtg 34140 ataccgaatg aatttttaaa acattcaggc aaggtgcatg ctcaggcatt ctttacacaa 34200 aacgggagta ataatgttgt tgttgaacgt caatttagct tcaatattga aaatgattta 34260 gttagtgggt ttgatggtat aacaaagctt gtttatatca aatctattca agatactatc 34320 gaagcagtcg gtaaagactt taaccaatta aagcaagata tggatgatac acaaacgtta 34380 atagcaaaag tgaatgatag tgcgacaaaa ggcattcaac aaatcgaaat caagcaaaac 34440 gaagctatac aagctattac tgcgacgcaa actagtgcaa cacaagctgt tacagctgaa 34500 gtcgataaaa tagttgaaaa agagcaagcg atttttgaac gtgttaacga agttgaacaa 34560 caaatcaatg gcgctgacct tgttaaaggt aattcaacaa caaattggca aaagtctaaa 34620 cttacagatg attacggtaa agcaattgaa tcgtatgagc agtccataga tagcgtttta 34680 agcgcagtta acacatctag gattattcat attactaatg caacagatgc gccagaaaag 34740 acggatatag gcacgttaga gaagcctgga caagatggtg ttgatgacgg ttcttcgttc 34800 gatgaatcaa cttatacatc aagcaaatct ggtgtgttag ttgtttatgt tgttgataat 34860 aatactgctc gtgcaacatg gtacccagac gattcaaacg atgagtacac aaaatacaaa 34920 atctacggca catggtaccc gttttataaa aagaatgatg gaaacttaac taagcaattt 34980 gttgaagaaa cgtctaacaa cgctttaaat caagctaagc agtatgtaga tgataaattc 35040 ggaacaacga gctggcaaca acataagatg acagaggcga atggtcaatc aattcaagtt 35100 aacttaaata atgcgcaagg cgatttggga tatttaactg ctggtaatta ctatgcaaca 35160 agagtgccgg atttaccagg tagtgttgaa agttatgagg gttatttatc ggtattcgtt 35220 aaagacgata caaacaagct atttaacttc acgccttata actctaaaaa gatttacaca 35280 cgatcaatca caaacggcag acttgagcaa cagtggacag ttcctaatga acataagtca 35340 acggtattgt tcgacggtgg agcaaatggt gtaggtacaa caatcaatct aaccgaacca 35400 tacacaaact attctatttt attagtaagt ggaacttatc caggtggcgt tattgaggga 35460 ttcggactaa ccacattacc taatgcaatt caattaagta aagcgaatgt agttgactca 35520 gacggtaacg gtggcggtat ttatgagtgt ttactatcca aaacaagtag cactacttta 35580 agaatcgata acgatgtgta ctttgattta ggtaaaacat caggttctgg agcgaatgcc 35640 aacaaagtta ctataactaa aattatgggg tggaaataat gaaaatcaca gtaaatgata 35700 aaaatgaagt tatcggatac gttaatactg gcggtttacg caatagttta gatgtagacg 35760 ataacaatgt gtctatcaaa ttcaaagaag agttcgaacc tagaaagttc gttttcacta 35820 acggcgaaat taaatacaat agcaatttcg aaaaagaaga cgtaccgaat gcatcaaacc 35880 aacaaagtgc gtcagattta agtgatgagg aacttcgcgg aatggttgca agtatgcaaa 35940 tgcagatgac gcaagtgaac atgttgacaa tgcaattgac gcaacaaaac gctatgttaa 36000 cacaacagtt gaccgaactg aaaactaaca aaacaaatac tgagggggac gtttaaatga 36060 tgaagatgat ttatccaact tttaaagaca ttaaaacttt ttatgtgtgg ggttgctata 36120 aaaatgagca aattaagtgg tacgtagaca tgggtgtaat cgacaaagaa gaatatgcat 36180 tgatcactgg tgaaaaatat ccagaggcaa aagatgaaaa gtcacaggtg taatgcttga 36240 ggctttttaa tttaacacaa agtaggtggc gtaatgtttg gatttaccaa acggcacgaa 36300 catgaatggc gaattagaag attagaagag aatgataaaa caatgcttag cactctcaat 36360 gagattaaat taggtcaaaa aactcaagag caagttaaca ttaaattaga taaaacttta 36420 gatgctatcc agagggaaag acagatagac gaaaaaaata agaaagaaaa cgacaaaaat 36480 atacgcgata tgaaaatgtg gattctcggt ttgataggga ctatcttcag tacgattgtc 36540 atagctttac taagaactat ttttggtatt taaaggaggt gattaccatg cttaaaggga 36600 ttttaggata tagcttctgg gcgtgcttct ggtttggtaa atgtaaataa cagttaagag 36660 tcagtgcttc ggcactggct ttttattttg attgaaatga ggtgcataca tgggattacc 36720 taacccaaag actagaaagc ctacagctag tgaagtggtg gagtgggcaa agtcgaatat 36780 tggtaagagg attaatatag ataattatcg gggcagtcaa tgttgggata cacctaactt 36840 tatttttaaa agatattggg gttttgtaac atggggcaat gctaaggata tggctaatta 36900 cagatatcct aagggtttcc gattctatcg ttattcatct ggatttgtac cggaacctgg 36960 agacatcgca gtttggcacc ctggcaacgg aataggttcg gacggacaca ccgcaatagt 37020 agtaggacca tctaataaaa gttattttta tagcgttgac caaaactggg ttaattctaa 37080 tagttggaca ggttctccag gaagattagt aagacaccct tatgtaagtg ttacaggctt 37140 tgttaggcct ccatactcaa aagatactag caaacctagt agtactgata caagttcagc 37200 atcaaaagcc aatgactcaa caattactgg cgaagcgaag aaaccgcaat ttaaagaagt 37260 taaaacagta aaatacactg cttacagcaa tgttttagat aaagaagagc acttcattga 37320 tcatatagtt gtaatgggtg atgaacgctc agatattcaa ggattatata taaaagaatc 37380 aatgcatatg cgttctgtag acgaactgta tacgcaaaga aataagttta taagcgatta 37440 tgaaataccg catttatatg tcgatagaga ggctacatgg cttgctagac caaccaattt 37500 tgatgacccg cgtcacccta attggctagt tattgaagta tgtggtggtc aaacagatag 37560 caaacgacaa ttcttattga atcaaataca agcgttaata cgtggtgttt ggttattgtc 37620 agggattgat aaaaacttat ctgaaacgac gttaaaggta gaccctaata tttggcgtag 37680 tatgaaagat ttaattaatt acgacttgat taagcaaggt ataccggata acgcaaagta 37740 tgagcaagtt aaaaagaaaa tgcttgagac atacattaaa cgagatatat tgacacgaga 37800 aaatataaaa gaagtaacga caaaaacaac aataagaatt agtgataaaa catcagttga 37860 cagtgcgtcc acacgaggcc ctactccatc agacgaaaaa ccaagcatcg ttactgaaac 37920 aagtccattc acattccagc aagcactgga tagacaaatg tctaggggta acccgaaaaa 37980 atctcataca tggggctggg ctaatgcaac acgagcacaa acgagctcgg caatgaatgt 38040 taagcgaata tgggaaagta acacgcaatg ctatcaaatg cttaatttag gcaagtatca 38100 aggcatttca gttagtgcgc ttaacaaaat acttaaagga aaaggaacgc tcgacggaca 38160 aggcaaagca ttcgcggaag cttgtaagaa aaacaacatt aacgaaattt atttgatcgc 38220 gcacgctttc ttagaaagtg gatacggaac aagtaacttc gctagtggta gatacggtgc 38280 atataattac ttcggtattg gtgcattcga caacgaccct gattatgcaa tgacgtttgc 38340 taaaaataaa ggttggacat ctccagcaaa agcaatcatg ggcggtgcta gcttcgtaag 38400 aaaggattac atcaataaag gtcaaaacac attgtaccga attagatgga atcctaagaa 38460 tccagctacc caccaatacg ctactgctat agagtggtgc caacatcaag caagtacaat 38520 cgctaagtta tataaacaaa tcggcttaaa aggtatctac ttcacaaggg ataaatataa 38580 ataaagaggt gtgtaaatgt acaaaataaa agatgttgaa acgagaataa aaaatgatgg 38640 tgttgactta ggtgacattg gctgtcgatt ttacactgaa gatgaaaata cagcatctat 38700 aagaataggt atcaatgaca aacaaggtcg tatcgatcta aaagcacatg gcttaacacc 38760 tagattacat ttgtttatgg aagatggctc tatattcaaa aatgagcccc ttattatcga 38820 cgatgttgta aaagggttcc ttacctacaa aatacctaaa aaggttatca aacacgctgg 38880 ttatgttcgc tgtaagctgt ttttagagaa agaagaagaa aaaatacatg tcgcaaactt 38940 ttctttcaat atcgttgata gtggtattga atctgctgta gcaaaagaaa tcgatgttaa 39000 attggtagat gatgctatta cgagaatttt aaaagataac gcgacagatt tattgagcaa 39060 agactttaaa gagaaaatag ataaagatgt catttcttac atcgaaaaga atgaaagtag 39120 atttaaaggt gcgaaaggtg ataaaggcga accgggacaa cctggtgcga aaggtgatac 39180 aggtaaaaaa ggagaacaag gcgcacccgg taaaaacggt actgtagtat caatcaatcc 39240 tgacactaaa atgtggcaaa ttgatggtaa agatacagat atcaaagcag aacctgagtt 39300 attggacaaa atcaatatcg caaatgttga agggttagaa gataaattgc aagaagttaa 39360 aaaaatcaaa gatacaactc tcaacgactc taaaacgtat acggattcaa aaattgctga 39420 actagttgat agcgcgcctg aatctatgaa tacattaaga gaattagcag aagcaataca 39480 aaacaactct atttcagaaa gtgtattgca acagattggc tcaaaagtta gtacagaaga 39540 ttttgaggaa ttcaaacaaa cactaaacga tttatatgct ccaaaaaatc ataatcatga 39600 tgagcggtat gttttgtcat ctcaagcttt tactaaacaa caagcggata atttatatca 39660 actaaaaagc gcatctcaac cgacggttaa aatttggaca ggaacagaaa atgaatataa 39720 ctatatatat caaaaagacc ctaatacact ttacttaatt aaggggtgat ttttatggaa 39780 ggtaatttta aaaatgtaaa gaagtttatt tacgaaggtg aagaatatac aaaagtatat 39840 gctggaaata tccaagtatg gaaaaagcct tcatcttttg taataaaacc cttacctaaa 39900 aataaatatc cggatagcat agaagaatca acagcaaaat ggacaataaa tggagttgaa 39960 cctaataaaa gttatcaggt gacaatagaa aatgtacgta gcggtataat gagggtttcg 40020 caaactaatt taggttcaag tgatttagga atatcaggag tcaatagcgg agttgcaagt 40080 aaaaatatca actttagtaa tccttcaggg atgttgtatg tcactataag tgatgtttat 40140 tcaggatctc caacattgac cattgaataa ttttaaacga ctaatttttt agtcgttttt 40200 tattttggat aaaaggagca aacaaatgga tgcaaaagta ataacaagat acatcgtatt 40260 gatcttagca ttagtaaatc aattcttagc gaacaaaggt attagcccga ttccagtaga 40320 cgatgagact atatcatcaa taatacttac tgttgttgct ttatatacta cgtataaaga 40380 caatccaaca tctcaagaag gtaaatgggc aaatcaaaag ctaaagaaat ataaagctga 40440 aaacaagtat agaaaagcaa cagggcaagc gccaattaaa gaagtaatga cacctacgaa 40500 tatgaacgac acaaatgatt tagggtaggt gttgaccaat gttgataaca aaaaaccaag 40560 cagaaaaatg gtttgataat tcattaggga agcagttcaa tcctgatttg ttttatggat 40620 ttcagtgtta cgattacgca aatatgtttt ttatgatagc aacaggcgaa aggttacaag 40680 gtttatacgc ttataatatt ccatttgata ataaagcaag gattgaaaaa tacgggcaaa 40740 taattaaaaa ctatgatagc tttttaccgc aaaagttgga tattgtcgtt ttcccgtcaa 40800 agtatggtgg cggagctgga catgttgaaa ttgttgagag cgcaaattta aacactttca 40860 catcatatgg gcaaaattgg aatggtaaag gttggacaaa tggcgttgcg caacctggtt 40920 ggggtcctga aactgttaca agacatgttc attattacga tgacccaatg tattttatta 40980 gattaaattt cccagataaa gtaagtgttg gagataaagc taaaagcgtt attaagcaag 41040 caactgccaa aaagcaagca gtaattaaac ctaaaaaaat tatgcttgta gccggtcatg 41100 gttataacga tcctggagca gtaggaaacg gaacaaacga acgcgatttt atccgtaaat 41160 atataacgcc aaatatcgct aagtatttaa gacatgcagg tcatgaagtt gcattatatg 41220 gtggctcaag tcaatcacaa gacatgtatc aagatactgc atacggtgtt aatgtaggaa 41280 ataataaaga ttatggatta tattgggtta aatcacaggg gtatgacatt gttctagaga 41340 ttcatttaga cgcagcagga gaaaatgcaa gtggtgggca tgttattatc tcaagtcaat 41400 tcaatgcgga tactattgat aaaagtatac aagatgttat taaaaataac ttaggacaaa 41460 taagaggtgt aacacctcgt aatgatttac tgaacgttaa tgtatcagca gaaataaata 41520 tcaattatcg tttatctgaa ttaggtttta ttactaataa aaaagatatg gattggatta 41580 agaagaatta tgacttgtat tctaaattaa tagctggtgc gattcatggt aagcctatag 41640 gtggtttggt agctggtaat gttaaaacat cagctaaaaa ccaaaaaaat ccaccagtgc 41700 cagcaggtta tacacttgat aagaataatg tgccttataa aaaagagact ggtaattaca 41760 cagttgccaa tgttaaaggt aataacgtaa gggacggcta ttcaactaat tcaagaatta 41820 caggtgtatt acctaataac gcaacaatca aatatgacgg cgcatattgc atcaatgggt 41880 atagatggat tacttatatt gctaatagtg gacaacgtcg ctatattgcg acaggagagg 41940 tagataaagc aggtaatagg ataagtagtt ttggtaagtt tagcacgatt tagtatttac 42000 ttagaataaa aattttgcta cattaattat agggaatctt acagttatta aataactatt 42060 tggatggatg ttaatattcc tatacacttt ttaacattac tctcaagatt taaatgtaga 42120 taacaggcag gtactacggt acttgcctat ttttttgtta taatgtaatt acattaccag 42180 taaccaatct ggcttaaaac cacatttccg gtagccaatc cggctatgca gaggacttac 42240 ttgcgtaaag tagtaagaag ctgactgcat atttaaacca cccatactag ttgctgggtg 42300 gttgtttttt atgttatatt ataaatgatc aaaccacacc acctattaat ttaggagtgt 42360 ggttattttt tatgcaaaaa aaacgaaaaa aagttcataa aaagtattgc atatcacgtt 42420 taaccgtgtt ataataaggt ataccagttg agaggaggat aaaaagtgtt agaaaatttt 42480 aaaactatag cagaaatcgc cttttataca atgtcagcaa ttgccatagc gaaaacattg 42540 aaaaaagacg ataagtaagt agacaagccc gaaagggctg tctatatata aattctaaca 42600 ctaaaatact atgaaaacaa tttacattat tttaatcatt cttatttgga taaacgtgtt 42660 tttaggcaac gatataagta aaagtgttgt tgcactgctt actactttac tgcttatcaa 42720 tttatggaag agggataaaa atgacagcaa taaaagaaat aattgaatca atagaaaagt 42780 tattcgaaaa agaaacggga tataaaattg ctaaaaattc cggattacca tatcaaactg 42840 tgcaagattt aagaaatgga aaaacatctt tatcagatgc cagatttaga acgataataa 42900 agttatacga gtatcaaaga tcgcttgaaa acgaagaaga taaataaaag gagccaaaaa 42960 tatgtttgtt acaaaagaag aatttaaaac tttgaatgta aaagaagtat ttgaatcagg 43020 taaaaacttt ataaaaatta cagatggaag acatgcaata tattgggtaa atgatagata 43080 cgtagtactt gaccataaaa aaggcgattt gtacccgcaa aaagcatacc caaaatatat 43140 caaaagaaaa ttagtaagtt aaataattag aaaaccacgt cttaattgac gtggttattt 43200 tttaggtttg cgcgtgtcaa atacgtgtca atttagttct atttctttag ttttctttct 43260 aaacttaatt gcttgtaaac cgcatagtta taggcttttc agctatatac caagataaga 43320 tttatcccgc cgtctccata aaaatatgct tggaaacctt gatttaatgg ggttttaatc 43380 tagcaagtgt caaatatgtg tcaagaaaat aattttctga cacgttgacc ttgctctttt 43440 ttatgttcat caagtaagtg agagtaggtg tctaaagtta tagatatatt ataatggcct 43500 aatcttttgc taatatattc aataggcata gttataggct tttcagctat ataccaagat 43560 aagatttatc ccgccg 43576 20 363 DNA Staphylococcus bacteriophage CDS (1)..(360) 20 atg gca ata tta gaa ggt att ttt gaa gaa tta aaa cta tta aat aag 48 Met Ala Ile Leu Glu Gly Ile Phe Glu Glu Leu Lys Leu Leu Asn Lys 1 5 10 15 aat tta cgt gtg cta aat act gaa cta tca act gta gat tca tca att 96 Asn Leu Arg Val Leu Asn Thr Glu Leu Ser Thr Val Asp Ser Ser Ile 20 25 30 gta caa gag aaa gtt aaa gaa gca cca atg cca aaa gat gaa aca gct 144 Val Gln Glu Lys Val Lys Glu Ala Pro Met Pro Lys Asp Glu Thr Ala 35 40 45 caa ctg gaa tca gtt gaa gaa gtt aag gaa act tct gct gat tta act 192 Gln Leu Glu Ser Val Glu Glu Val Lys Glu Thr Ser Ala Asp Leu Thr 50 55 60 aaa gat tat gtt tta tca gta gga aaa gag ttc ctt aaa aaa gca gat 240 Lys Asp Tyr Val Leu Ser Val Gly Lys Glu Phe Leu Lys Lys Ala Asp 65 70 75 80 act tct gat aag aaa gaa ttt aga aat aaa ctt aac gaa ctt ggt gcg 288 Thr Ser Asp Lys Lys Glu Phe Arg Asn Lys Leu Asn Glu Leu Gly Ala 85 90 95 gat aag cta tct act atc aaa gaa gag cat tat gaa aaa att gtt gat 336 Asp Lys Leu Ser Thr Ile Lys Glu Glu His Tyr Glu Lys Ile Val Asp 100 105 110 ttt atg aat gcg aga ata aat gca tga 363 Phe Met Asn Ala Arg Ile Asn Ala 115 120 21 120 PRT Staphylococcus bacteriophage 21 Met Ala Ile Leu Glu Gly Ile Phe Glu Glu Leu Lys Leu Leu Asn Lys 1 5 10 15 Asn Leu Arg Val Leu Asn Thr Glu Leu Ser Thr Val Asp Ser Ser Ile 20 25 30 Val Gln Glu Lys Val Lys Glu Ala Pro Met Pro Lys Asp Glu Thr Ala 35 40 45 Gln Leu Glu Ser Val Glu Glu Val Lys Glu Thr Ser Ala Asp Leu Thr 50 55 60 Lys Asp Tyr Val Leu Ser Val Gly Lys Glu Phe Leu Lys Lys Ala Asp 65 70 75 80 Thr Ser Asp Lys Lys Glu Phe Arg Asn Lys Leu Asn Glu Leu Gly Ala 85 90 95 Asp Lys Leu Ser Thr Ile Lys Glu Glu His Tyr Glu Lys Ile Val Asp 100 105 110 Phe Met Asn Ala Arg Ile Asn Ala 115 120 22 300 DNA Staphylococcus bacteriophage CDS (1)..(297) 22 atg ttt gga ttt acc aaa cga cac gaa caa gat tgg cgt tta acg cga 48 Met Phe Gly Phe Thr Lys Arg His Glu Gln Asp Trp Arg Leu Thr Arg 1 5 10 15 tta gaa gaa aat gat aag act atg ttt gaa aaa ttc gac aga ata gaa 96 Leu Glu Glu Asn Asp Lys Thr Met Phe Glu Lys Phe Asp Arg Ile Glu 20 25 30 gac agt ctg aga acg caa gaa aaa att tat gac aag tta gat aga aat 144 Asp Ser Leu Arg Thr Gln Glu Lys Ile Tyr Asp Lys Leu Asp Arg Asn 35 40 45 ttc gaa gaa cta agg cgt gac aaa gaa gaa gat gaa aaa aat aaa gag 192 Phe Glu Glu Leu Arg Arg Asp Lys Glu Glu Asp Glu Lys Asn Lys Glu 50 55 60 aaa aat gct aaa aat att aga gac atc aag atg tgg att cta gga tta 240 Lys Asn Ala Lys Asn Ile Arg Asp Ile Lys Met Trp Ile Leu Gly Leu 65 70 75 80 ata ggg acg att cta agt aca ttt gtt ata gcc ttg tta aaa act att 288 Ile Gly Thr Ile Leu Ser Thr Phe Val Ile Ala Leu Leu Lys Thr Ile 85 90 95 ttt ggc att taa 300 Phe Gly Ile 23 99 PRT Staphylococcus bacteriophage 23 Met Phe Gly Phe Thr Lys Arg His Glu Gln Asp Trp Arg Leu Thr Arg 1 5 10 15 Leu Glu Glu Asn Asp Lys Thr Met Phe Glu Lys Phe Asp Arg Ile Glu 20 25 30 Asp Ser Leu Arg Thr Gln Glu Lys Ile Tyr Asp Lys Leu Asp Arg Asn 35 40 45 Phe Glu Glu Leu Arg Arg Asp Lys Glu Glu Asp Glu Lys Asn Lys Glu 50 55 60 Lys Asn Ala Lys Asn Ile Arg Asp Ile Lys Met Trp Ile Leu Gly Leu 65 70 75 80 Ile Gly Thr Ile Leu Ser Thr Phe Val Ile Ala Leu Leu Lys Thr Ile 85 90 95 Phe Gly Ile 24 186 DNA Staphylococcus bacteriophage CDS (1)..(183) 24 atg caa cat caa gct tat atc aat gct tct gtt gac att aga att cct 48 Met Gln His Gln Ala Tyr Ile Asn Ala Ser Val Asp Ile Arg Ile Pro 1 5 10 15 aca gaa gtc gaa agt gtt aat tac aat cag att gat aaa gaa aaa gaa 96 Thr Glu Val Glu Ser Val Asn Tyr Asn Gln Ile Asp Lys Glu Lys Glu 20 25 30 aat ttg gcg gac tat tta ttt aat aat cca ggt gaa cta tta aaa tat 144 Asn Leu Ala Asp Tyr Leu Phe Asn Asn Pro Gly Glu Leu Leu Lys Tyr 35 40 45 aac gtt ata aat att aag gtt tta gat tta gag gtg gaa tga 186 Asn Val Ile Asn Ile Lys Val Leu Asp Leu Glu Val Glu 50 55 60 25 61 PRT Staphylococcus bacteriophage 25 Met Gln His Gln Ala Tyr Ile Asn Ala Ser Val Asp Ile Arg Ile Pro 1 5 10 15 Thr Glu Val Glu Ser Val Asn Tyr Asn Gln Ile Asp Lys Glu Lys Glu 20 25 30 Asn Leu Ala Asp Tyr Leu Phe Asn Asn Pro Gly Glu Leu Leu Lys Tyr 35 40 45 Asn Val Ile Asn Ile Lys Val Leu Asp Leu Glu Val Glu 50 55 60 26 4530 DNA Staphylococcus bacteriophage CDS (1)..(4527) 26 atg gga gaa aga ata aaa ggt tta tct ata ggt ttg gat tta gat gca 48 Met Gly Glu Arg Ile Lys Gly Leu Ser Ile Gly Leu Asp Leu Asp Ala 1 5 10 15 gca aat tta aat aga tca ttt gca gaa atc aaa cga aac ttt aaa act 96 Ala Asn Leu Asn Arg Ser Phe Ala Glu Ile Lys Arg Asn Phe Lys Thr 20 25 30 tta aat tct gac tta aaa tta aca ggc aac aac ttc aaa tat acc gaa 144 Leu Asn Ser Asp Leu Lys Leu Thr Gly Asn Asn Phe Lys Tyr Thr Glu 35 40 45 aaa tca act gat agt tac aaa caa agg att aaa gaa ctt gat gga act 192 Lys Ser Thr Asp Ser Tyr Lys Gln Arg Ile Lys Glu Leu Asp Gly Thr 50 55 60 atc aca ggt tat aag aaa aac gtt gat gat tta gcc aag caa tat gac 240 Ile Thr Gly Tyr Lys Lys Asn Val Asp Asp Leu Ala Lys Gln Tyr Asp 65 70 75 80 aag gta tct caa gaa cag ggc gaa aac agt gca gaa gct caa aag tta 288 Lys Val Ser Gln Glu Gln Gly Glu Asn Ser Ala Glu Ala Gln Lys Leu 85 90 95 cga caa gaa tat aac aaa caa gca aat gag ctg aat tat tta gaa aga 336 Arg Gln Glu Tyr Asn Lys Gln Ala Asn Glu Leu Asn Tyr Leu Glu Arg 100 105 110 gaa tta caa aaa aca tca gcc gaa ttt gaa gag ttc aaa aaa gct caa 384 Glu Leu Gln Lys Thr Ser Ala Glu Phe Glu Glu Phe Lys Lys Ala Gln 115 120 125 gtt gaa gct caa aga atg gca gaa agt ggc tgg gga aaa acc agt aaa 432 Val Glu Ala Gln Arg Met Ala Glu Ser Gly Trp Gly Lys Thr Ser Lys 130 135 140 gtt ttt gaa agt atg gga cct aaa tta aca aaa atg ggt gat ggt tta 480 Val Phe Glu Ser Met Gly Pro Lys Leu Thr Lys Met Gly Asp Gly Leu 145 150 155 160 aaa tcc att ggt aaa ggt ttg atg att ggt gta act gca cct gtt tta 528 Lys Ser Ile Gly Lys Gly Leu Met Ile Gly Val Thr Ala Pro Val Leu 165 170 175 ggt att gca gca gca tca gga aaa gct ttt gca gaa gtt gat aaa ggt 576 Gly Ile Ala Ala Ala Ser Gly Lys Ala Phe Ala Glu Val Asp Lys Gly 180 185 190 tta gat act gtt act caa gca aca ggc gca aca ggc agt gaa tta aaa 624 Leu Asp Thr Val Thr Gln Ala Thr Gly Ala Thr Gly Ser Glu Leu Lys 195 200 205 aaa ttg cag aac tca ttt aaa gat gtt tat ggc aat ttt cca gca gat 672 Lys Leu Gln Asn Ser Phe Lys Asp Val Tyr Gly Asn Phe Pro Ala Asp 210 215 220 gct gaa act gtt ggt gga gtt tta gga gaa gtt aat aca agg tta ggt 720 Ala Glu Thr Val Gly Gly Val Leu Gly Glu Val Asn Thr Arg Leu Gly 225 230 235 240 ttt aca ggt aaa gaa ctt gaa aat gcc aca gag tca ttc ttg aaa ttc 768 Phe Thr Gly Lys Glu Leu Glu Asn Ala Thr Glu Ser Phe Leu Lys Phe 245 250 255 agt cat ata aca ggt tct gac ggt gtg caa gcc gta cag tta att acc 816 Ser His Ile Thr Gly Ser Asp Gly Val Gln Ala Val Gln Leu Ile Thr 260 265 270 cgt gca atg ggc gat gca ggt atc gaa gca agt gaa tat caa agt gtt 864 Arg Ala Met Gly Asp Ala Gly Ile Glu Ala Ser Glu Tyr Gln Ser Val 275 280 285 ttg gat atg gta gca aaa gcg gcg caa gct agt ggg ata agt gtt gat 912 Leu Asp Met Val Ala Lys Ala Ala Gln Ala Ser Gly Ile Ser Val Asp 290 295 300 aca tta gct gat agt att act aaa tac ggc gct cca atg aga gct atg 960 Thr Leu Ala Asp Ser Ile Thr Lys Tyr Gly Ala Pro Met Arg Ala Met 305 310 315 320 ggc ttt gag atg aaa gaa tca att gct tta ttc tct caa tgg gaa aag 1008 Gly Phe Glu Met Lys Glu Ser Ile Ala Leu Phe Ser Gln Trp Glu Lys 325 330 335 tca ggc gtt aat act gaa ata gca ttc agt ggt ttg aaa aaa gct ata 1056 Ser Gly Val Asn Thr Glu Ile Ala Phe Ser Gly Leu Lys Lys Ala Ile 340 345 350 tca aat tgg ggt aaa gct ggt aaa aac cca aga gaa gaa ttt aag aag 1104 Ser Asn Trp Gly Lys Ala Gly Lys Asn Pro Arg Glu Glu Phe Lys Lys 355 360 365 aca tta gca gaa att gaa aag acg ccg gat ata gct agc gca aca agt 1152 Thr Leu Ala Glu Ile Glu Lys Thr Pro Asp Ile Ala Ser Ala Thr Ser 370 375 380 tta gcg att gaa gca ttt ggt gca aag gca ggt cct gat tta gca gac 1200 Leu Ala Ile Glu Ala Phe Gly Ala Lys Ala Gly Pro Asp Leu Ala Asp 385 390 395 400 gct att aaa ggt ggt cgc ttt agt tat caa gaa ttt tta aaa act att 1248 Ala Ile Lys Gly Gly Arg Phe Ser Tyr Gln Glu Phe Leu Lys Thr Ile 405 410 415 gaa gat tcc caa ggc aca gta aac caa aca ttt aaa gat tct gaa agt 1296 Glu Asp Ser Gln Gly Thr Val Asn Gln Thr Phe Lys Asp Ser Glu Ser 420 425 430 ggc tcc gaa aga ttt aaa gta gca atg aat aaa tta aaa tta gta ggt 1344 Gly Ser Glu Arg Phe Lys Val Ala Met Asn Lys Leu Lys Leu Val Gly 435 440 445 gct gat gta tgg gct tct att gaa agt gcg ttt gct ccc gta atg gaa 1392 Ala Asp Val Trp Ala Ser Ile Glu Ser Ala Phe Ala Pro Val Met Glu 450 455 460 gaa tta atc aaa aag cta tct ata gcg gtt gat tgg ttt tcc aat tta 1440 Glu Leu Ile Lys Lys Leu Ser Ile Ala Val Asp Trp Phe Ser Asn Leu 465 470 475 480 agt gat ggt tct aaa aga tca att gtt att ttc agt ggt att gct gct 1488 Ser Asp Gly Ser Lys Arg Ser Ile Val Ile Phe Ser Gly Ile Ala Ala 485 490 495 gca att ggt cct gta gtt ttt ggg tta ggt gca ttt ata agt aca att 1536 Ala Ile Gly Pro Val Val Phe Gly Leu Gly Ala Phe Ile Ser Thr Ile 500 505 510 ggc aat gca gta act gta tta gct cca ttg tta gct agt att gca aag 1584 Gly Asn Ala Val Thr Val Leu Ala Pro Leu Leu Ala Ser Ile Ala Lys 515 520 525 gct ggt gga ttg att agt ttt tta tcg act aaa gta cct ata tta gga 1632 Ala Gly Gly Leu Ile Ser Phe Leu Ser Thr Lys Val Pro Ile Leu Gly 530 535 540 act gtc ttc aca gct tta act ggt cca att ggc att gta tta ggt gta 1680 Thr Val Phe Thr Ala Leu Thr Gly Pro Ile Gly Ile Val Leu Gly Val 545 550 555 560 ttg gct ggt tta gca gtc gca ttt aca att gct tat aag aaa tct gaa 1728 Leu Ala Gly Leu Ala Val Ala Phe Thr Ile Ala Tyr Lys Lys Ser Glu 565 570 575 aca ttt aga aat ttt gtt aat ggt gca att gaa agt gtt aaa caa aca 1776 Thr Phe Arg Asn Phe Val Asn Gly Ala Ile Glu Ser Val Lys Gln Thr 580 585 590 ttt agt aat ttt att caa ttt att caa cct ttc gtt gat tct gtt aaa 1824 Phe Ser Asn Phe Ile Gln Phe Ile Gln Pro Phe Val Asp Ser Val Lys 595 600 605 aac atc ttt aaa caa gcg ata tca gca ata gtt gat ttc gca aaa gat 1872 Asn Ile Phe Lys Gln Ala Ile Ser Ala Ile Val Asp Phe Ala Lys Asp 610 615 620 att tgg agt caa atc aat gga ttc ttt aat gaa aac gga att tcc att 1920 Ile Trp Ser Gln Ile Asn Gly Phe Phe Asn Glu Asn Gly Ile Ser Ile 625 630 635 640 gtt caa gca ctt caa aat ata tgc aac ttt att aaa gcg ata ttt gaa 1968 Val Gln Ala Leu Gln Asn Ile Cys Asn Phe Ile Lys Ala Ile Phe Glu 645 650 655 ttt att tta aat ttt gta att aaa cca att atg ttc gcg att tgg caa 2016 Phe Ile Leu Asn Phe Val Ile Lys Pro Ile Met Phe Ala Ile Trp Gln 660 665 670 gtg atg caa ttt att tgg ccg gcg gtt aaa gcc ttg att gtc agt act 2064 Val Met Gln Phe Ile Trp Pro Ala Val Lys Ala Leu Ile Val Ser Thr 675 680 685 tgg gag aac ata aaa ggt gta ata caa ggt gct tta aat atc ata ctt 2112 Trp Glu Asn Ile Lys Gly Val Ile Gln Gly Ala Leu Asn Ile Ile Leu 690 695 700 ggc ttg att aag ttc ttc tca agt tta ttc gtt ggt gat tgg cga gga 2160 Gly Leu Ile Lys Phe Phe Ser Ser Leu Phe Val Gly Asp Trp Arg Gly 705 710 715 720 gtt tgg gac gcc gtt gtg atg att ctt aaa gga gca gtt caa tta att 2208 Val Trp Asp Ala Val Val Met Ile Leu Lys Gly Ala Val Gln Leu Ile 725 730 735 tgg aat tta gtt caa tta tgg ttt gta ggt aaa ata ctt ggt gtt gtt 2256 Trp Asn Leu Val Gln Leu Trp Phe Val Gly Lys Ile Leu Gly Val Val 740 745 750 agg tac ttt ggc ggg ttg cta aaa gga ttg ata gca gga att tgg gac 2304 Arg Tyr Phe Gly Gly Leu Leu Lys Gly Leu Ile Ala Gly Ile Trp Asp 755 760 765 gta ata aga agt ata ttc agt aaa tct tta tca gca att tgg aat gca 2352 Val Ile Arg Ser Ile Phe Ser Lys Ser Leu Ser Ala Ile Trp Asn Ala 770 775 780 aca aaa agt att ttt gga ttt tta ttt aat agc gta aaa tca att ttc 2400 Thr Lys Ser Ile Phe Gly Phe Leu Phe Asn Ser Val Lys Ser Ile Phe 785 790 795 800 aca aat atg aaa aat tgg tta tct aat act tgg agc agt atc cgt acg 2448 Thr Asn Met Lys Asn Trp Leu Ser Asn Thr Trp Ser Ser Ile Arg Thr 805 810 815 aat aca ata gga aaa gcg cag tca tta ttt agt ggc gtc aaa tca aaa 2496 Asn Thr Ile Gly Lys Ala Gln Ser Leu Phe Ser Gly Val Lys Ser Lys 820 825 830 ttt act aat tta tgg aat gcg acg aaa gaa att ttt agt aat tta aga 2544 Phe Thr Asn Leu Trp Asn Ala Thr Lys Glu Ile Phe Ser Asn Leu Arg 835 840 845 aat tgg atg tca aat att tgg aat tcc att aaa gat aat acg gta gga 2592 Asn Trp Met Ser Asn Ile Trp Asn Ser Ile Lys Asp Asn Thr Val Gly 850 855 860 att gca agc cgt tta tgg agt aag gta cgt gga att ttc aca aat atg 2640 Ile Ala Ser Arg Leu Trp Ser Lys Val Arg Gly Ile Phe Thr Asn Met 865 870 875 880 cgc gat ggc ttg agt tcc att ata gat aag att aaa agt cat atc ggc 2688 Arg Asp Gly Leu Ser Ser Ile Ile Asp Lys Ile Lys Ser His Ile Gly 885 890 895 ggt atg gta agc gct att aaa aaa gga ctt aat aaa tta atc gac ggt 2736 Gly Met Val Ser Ala Ile Lys Lys Gly Leu Asn Lys Leu Ile Asp Gly 900 905 910 tta aac tgg gtc ggt ggt aag ttg gga atg gat aaa ata cct aag tta 2784 Leu Asn Trp Val Gly Gly Lys Leu Gly Met Asp Lys Ile Pro Lys Leu 915 920 925 cac act ggt aca gag cac aca cat act act aca aga tta gtt aag aac 2832 His Thr Gly Thr Glu His Thr His Thr Thr Thr Arg Leu Val Lys Asn 930 935 940 ggt aag att gca cgt gac aca ttc gct aca gtt ggg gat aag gga cgc 2880 Gly Lys Ile Ala Arg Asp Thr Phe Ala Thr Val Gly Asp Lys Gly Arg 945 950 955 960 gga aat ggt cca aat ggt ttt aga aat gaa atg att gaa ttc cct aac 2928 Gly Asn Gly Pro Asn Gly Phe Arg Asn Glu Met Ile Glu Phe Pro Asn 965 970 975 ggt aaa cgt gta atc aca cct aat aca gat act acc gct tat tta cct 2976 Gly Lys Arg Val Ile Thr Pro Asn Thr Asp Thr Thr Ala Tyr Leu Pro 980 985 990 aaa ggc tca aaa gta tac aac ggt gca caa act tat tca atg tta aac 3024 Lys Gly Ser Lys Val Tyr Asn Gly Ala Gln Thr Tyr Ser Met Leu Asn 995 1000 1005 gga acg ctt cca aga ttt agt tta ggt act atg tgg aaa gat att aaa 3072 Gly Thr Leu Pro Arg Phe Ser Leu Gly Thr Met Trp Lys Asp Ile Lys 1010 1015 1020 tct ggt gca tca tcg gca ttt aac tgg aca aaa gat aaa ata ggt aaa 3120 Ser Gly Ala Ser Ser Ala Phe Asn Trp Thr Lys Asp Lys Ile Gly Lys 1025 1030 1035 1040 ggt acc aaa tgg ctt ggc gat aaa gtt ggc gat gtt tta gat ttt atg 3168 Gly Thr Lys Trp Leu Gly Asp Lys Val Gly Asp Val Leu Asp Phe Met 1045 1050 1055 gaa aat cca ggc aaa ctt tta aat tat ata ctt gaa gct ttt gga att 3216 Glu Asn Pro Gly Lys Leu Leu Asn Tyr Ile Leu Glu Ala Phe Gly Ile 1060 1065 1070 gat ttc aat tct tta act aaa ggt atg gga att gca ggc gac ata aca 3264 Asp Phe Asn Ser Leu Thr Lys Gly Met Gly Ile Ala Gly Asp Ile Thr 1075 1080 1085 aaa gct gca tgg tct aag att aag aaa agt gct act gat tgg ata aaa 3312 Lys Ala Ala Trp Ser Lys Ile Lys Lys Ser Ala Thr Asp Trp Ile Lys 1090 1095 1100 gaa aat tta gaa gct atg ggc ggt ggc gat tta gtc ggc gga ata tta 3360 Glu Asn Leu Glu Ala Met Gly Gly Gly Asp Leu Val Gly Gly Ile Leu 1105 1110 1115 1120 gac cct gac aaa att aat tat cat tat gga cgt acc gca gct tat acc 3408 Asp Pro Asp Lys Ile Asn Tyr His Tyr Gly Arg Thr Ala Ala Tyr Thr 1125 1130 1135 gct gca act gga aga cca ttt cat gaa ggt gtc gat ttt cca ttt gta 3456 Ala Ala Thr Gly Arg Pro Phe His Glu Gly Val Asp Phe Pro Phe Val 1140 1145 1150 tat caa gaa gtt aga acg ccg atg ggt ggc aga ctt aca aga atg cca 3504 Tyr Gln Glu Val Arg Thr Pro Met Gly Gly Arg Leu Thr Arg Met Pro 1155 1160 1165 ttt atg tct ggt ggt tat ggt aat tat gta aaa att act agt ggc gtt 3552 Phe Met Ser Gly Gly Tyr Gly Asn Tyr Val Lys Ile Thr Ser Gly Val 1170 1175 1180 atc gat atg cta ttt gcg cat ttg aaa aac ttt agc aaa tca cca cct 3600 Ile Asp Met Leu Phe Ala His Leu Lys Asn Phe Ser Lys Ser Pro Pro 1185 1190 1195 1200 agt ggc acg atg gta aag ccc ggt gat gtt gtt ggt tta act ggt aat 3648 Ser Gly Thr Met Val Lys Pro Gly Asp Val Val Gly Leu Thr Gly Asn 1205 1210 1215 acc gga ttt agt aca gga cca cat tta cat ttt gaa atg agg aga aat 3696 Thr Gly Phe Ser Thr Gly Pro His Leu His Phe Glu Met Arg Arg Asn 1220 1225 1230 gga cga cat ttt gac cct gaa cca tat tta agg aat gct aag aaa aaa 3744 Gly Arg His Phe Asp Pro Glu Pro Tyr Leu Arg Asn Ala Lys Lys Lys 1235 1240 1245 gga aga tta tca ata ggt ggt ggc ggt gct act tct gga agt ggc gca 3792 Gly Arg Leu Ser Ile Gly Gly Gly Gly Ala Thr Ser Gly Ser Gly Ala 1250 1255 1260 act tat gcc agt cga gta atc cga caa gcg caa agt att tta ggt ggt 3840 Thr Tyr Ala Ser Arg Val Ile Arg Gln Ala Gln Ser Ile Leu Gly Gly 1265 1270 1275 1280 cgt tat aaa ggt aaa tgg att cat gac caa atg atg cgc gtt gca aaa 3888 Arg Tyr Lys Gly Lys Trp Ile His Asp Gln Met Met Arg Val Ala Lys 1285 1290 1295 cgt gaa agt aac tac cag tca aat gca gtg aat aac tgg gat ata aat 3936 Arg Glu Ser Asn Tyr Gln Ser Asn Ala Val Asn Asn Trp Asp Ile Asn 1300 1305 1310 gct caa aga gga gac cca tca aga gga tta ttc caa atc atc ggc tca 3984 Ala Gln Arg Gly Asp Pro Ser Arg Gly Leu Phe Gln Ile Ile Gly Ser 1315 1320 1325 act ttt aga gca aac gct aaa cgt gga tat act aac ttt aat aat cca 4032 Thr Phe Arg Ala Asn Ala Lys Arg Gly Tyr Thr Asn Phe Asn Asn Pro 1330 1335 1340 gta cat caa ggt atc tca gca atg cag tac att gtt aga cga tat ggt 4080 Val His Gln Gly Ile Ser Ala Met Gln Tyr Ile Val Arg Arg Tyr Gly 1345 1350 1355 1360 tgg ggt ggt ttt aaa cgt gct ggt gat tac gca tat gct aca ggt gga 4128 Trp Gly Gly Phe Lys Arg Ala Gly Asp Tyr Ala Tyr Ala Thr Gly Gly 1365 1370 1375 aaa gtt ttt gat ggt tgg tat aac tta ggt gaa gac ggt cat cca gaa 4176 Lys Val Phe Asp Gly Trp Tyr Asn Leu Gly Glu Asp Gly His Pro Glu 1380 1385 1390 tgg att att cca aca gat cca gct cgt aga aat gat gca atg aag att 4224 Trp Ile Ile Pro Thr Asp Pro Ala Arg Arg Asn Asp Ala Met Lys Ile 1395 1400 1405 ttg cat tat gca gca gca gaa gta aga ggg aaa aaa gcg agt aaa aat 4272 Leu His Tyr Ala Ala Ala Glu Val Arg Gly Lys Lys Ala Ser Lys Asn 1410 1415 1420 aag cgt cct agc caa tta tca gac tta aac ggg ttt gat gat cct agc 4320 Lys Arg Pro Ser Gln Leu Ser Asp Leu Asn Gly Phe Asp Asp Pro Ser 1425 1430 1435 1440 tta tta ttg aaa atg att gaa caa cag caa caa caa ata gct tta tta 4368 Leu Leu Leu Lys Met Ile Glu Gln Gln Gln Gln Gln Ile Ala Leu Leu 1445 1450 1455 ctg aaa ata gca caa tct aac gat gtg att gca gat aaa gat tat cag 4416 Leu Lys Ile Ala Gln Ser Asn Asp Val Ile Ala Asp Lys Asp Tyr Gln 1460 1465 1470 ccg att att gac gaa tac gct ttt gat aaa aag gtg aac gcg tct ata 4464 Pro Ile Ile Asp Glu Tyr Ala Phe Asp Lys Lys Val Asn Ala Ser Ile 1475 1480 1485 gaa aag cga gaa agg caa gaa tca aca aaa gta aag ttt aga aaa gga 4512 Glu Lys Arg Glu Arg Gln Glu Ser Thr Lys Val Lys Phe Arg Lys Gly 1490 1495 1500 gga att gct att caa tga 4530 Gly Ile Ala Ile Gln 1505 27 1509 PRT Staphylococcus bacteriophage 27 Met Gly Glu Arg Ile Lys Gly Leu Ser Ile Gly Leu Asp Leu Asp Ala 1 5 10 15 Ala Asn Leu Asn Arg Ser Phe Ala Glu Ile Lys Arg Asn Phe Lys Thr 20 25 30 Leu Asn Ser Asp Leu Lys Leu Thr Gly Asn Asn Phe Lys Tyr Thr Glu 35 40 45 Lys Ser Thr Asp Ser Tyr Lys Gln Arg Ile Lys Glu Leu Asp Gly Thr 50 55 60 Ile Thr Gly Tyr Lys Lys Asn Val Asp Asp Leu Ala Lys Gln Tyr Asp 65 70 75 80 Lys Val Ser Gln Glu Gln Gly Glu Asn Ser Ala Glu Ala Gln Lys Leu 85 90 95 Arg Gln Glu Tyr Asn Lys Gln Ala Asn Glu Leu Asn Tyr Leu Glu Arg 100 105 110 Glu Leu Gln Lys Thr Ser Ala Glu Phe Glu Glu Phe Lys Lys Ala Gln 115 120 125 Val Glu Ala Gln Arg Met Ala Glu Ser Gly Trp Gly Lys Thr Ser Lys 130 135 140 Val Phe Glu Ser Met Gly Pro Lys Leu Thr Lys Met Gly Asp Gly Leu 145 150 155 160 Lys Ser Ile Gly Lys Gly Leu Met Ile Gly Val Thr Ala Pro Val Leu 165 170 175 Gly Ile Ala Ala Ala Ser Gly Lys Ala Phe Ala Glu Val Asp Lys Gly 180 185 190 Leu Asp Thr Val Thr Gln Ala Thr Gly Ala Thr Gly Ser Glu Leu Lys 195 200 205 Lys Leu Gln Asn Ser Phe Lys Asp Val Tyr Gly Asn Phe Pro Ala Asp 210 215 220 Ala Glu Thr Val Gly Gly Val Leu Gly Glu Val Asn Thr Arg Leu Gly 225 230 235 240 Phe Thr Gly Lys Glu Leu Glu Asn Ala Thr Glu Ser Phe Leu Lys Phe 245 250 255 Ser His Ile Thr Gly Ser Asp Gly Val Gln Ala Val Gln Leu Ile Thr 260 265 270 Arg Ala Met Gly Asp Ala Gly Ile Glu Ala Ser Glu Tyr Gln Ser Val 275 280 285 Leu Asp Met Val Ala Lys Ala Ala Gln Ala Ser Gly Ile Ser Val Asp 290 295 300 Thr Leu Ala Asp Ser Ile Thr Lys Tyr Gly Ala Pro Met Arg Ala Met 305 310 315 320 Gly Phe Glu Met Lys Glu Ser Ile Ala Leu Phe Ser Gln Trp Glu Lys 325 330 335 Ser Gly Val Asn Thr Glu Ile Ala Phe Ser Gly Leu Lys Lys Ala Ile 340 345 350 Ser Asn Trp Gly Lys Ala Gly Lys Asn Pro Arg Glu Glu Phe Lys Lys 355 360 365 Thr Leu Ala Glu Ile Glu Lys Thr Pro Asp Ile Ala Ser Ala Thr Ser 370 375 380 Leu Ala Ile Glu Ala Phe Gly Ala Lys Ala Gly Pro Asp Leu Ala Asp 385 390 395 400 Ala Ile Lys Gly Gly Arg Phe Ser Tyr Gln Glu Phe Leu Lys Thr Ile 405 410 415 Glu Asp Ser Gln Gly Thr Val Asn Gln Thr Phe Lys Asp Ser Glu Ser 420 425 430 Gly Ser Glu Arg Phe Lys Val Ala Met Asn Lys Leu Lys Leu Val Gly 435 440 445 Ala Asp Val Trp Ala Ser Ile Glu Ser Ala Phe Ala Pro Val Met Glu 450 455 460 Glu Leu Ile Lys Lys Leu Ser Ile Ala Val Asp Trp Phe Ser Asn Leu 465 470 475 480 Ser Asp Gly Ser Lys Arg Ser Ile Val Ile Phe Ser Gly Ile Ala Ala 485 490 495 Ala Ile Gly Pro Val Val Phe Gly Leu Gly Ala Phe Ile Ser Thr Ile 500 505 510 Gly Asn Ala Val Thr Val Leu Ala Pro Leu Leu Ala Ser Ile Ala Lys 515 520 525 Ala Gly Gly Leu Ile Ser Phe Leu Ser Thr Lys Val Pro Ile Leu Gly 530 535 540 Thr Val Phe Thr Ala Leu Thr Gly Pro Ile Gly Ile Val Leu Gly Val 545 550 555 560 Leu Ala Gly Leu Ala Val Ala Phe Thr Ile Ala Tyr Lys Lys Ser Glu 565 570 575 Thr Phe Arg Asn Phe Val Asn Gly Ala Ile Glu Ser Val Lys Gln Thr 580 585 590 Phe Ser Asn Phe Ile Gln Phe Ile Gln Pro Phe Val Asp Ser Val Lys 595 600 605 Asn Ile Phe Lys Gln Ala Ile Ser Ala Ile Val Asp Phe Ala Lys Asp 610 615 620 Ile Trp Ser Gln Ile Asn Gly Phe Phe Asn Glu Asn Gly Ile Ser Ile 625 630 635 640 Val Gln Ala Leu Gln Asn Ile Cys Asn Phe Ile Lys Ala Ile Phe Glu 645 650 655 Phe Ile Leu Asn Phe Val Ile Lys Pro Ile Met Phe Ala Ile Trp Gln 660 665 670 Val Met Gln Phe Ile Trp Pro Ala Val Lys Ala Leu Ile Val Ser Thr 675 680 685 Trp Glu Asn Ile Lys Gly Val Ile Gln Gly Ala Leu Asn Ile Ile Leu 690 695 700 Gly Leu Ile Lys Phe Phe Ser Ser Leu Phe Val Gly Asp Trp Arg Gly 705 710 715 720 Val Trp Asp Ala Val Val Met Ile Leu Lys Gly Ala Val Gln Leu Ile 725 730 735 Trp Asn Leu Val Gln Leu Trp Phe Val Gly Lys Ile Leu Gly Val Val 740 745 750 Arg Tyr Phe Gly Gly Leu Leu Lys Gly Leu Ile Ala Gly Ile Trp Asp 755 760 765 Val Ile Arg Ser Ile Phe Ser Lys Ser Leu Ser Ala Ile Trp Asn Ala 770 775 780 Thr Lys Ser Ile Phe Gly Phe Leu Phe Asn Ser Val Lys Ser Ile Phe 785 790 795 800 Thr Asn Met Lys Asn Trp Leu Ser Asn Thr Trp Ser Ser Ile Arg Thr 805 810 815 Asn Thr Ile Gly Lys Ala Gln Ser Leu Phe Ser Gly Val Lys Ser Lys 820 825 830 Phe Thr Asn Leu Trp Asn Ala Thr Lys Glu Ile Phe Ser Asn Leu Arg 835 840 845 Asn Trp Met Ser Asn Ile Trp Asn Ser Ile Lys Asp Asn Thr Val Gly 850 855 860 Ile Ala Ser Arg Leu Trp Ser Lys Val Arg Gly Ile Phe Thr Asn Met 865 870 875 880 Arg Asp Gly Leu Ser Ser Ile Ile Asp Lys Ile Lys Ser His Ile Gly 885 890 895 Gly Met Val Ser Ala Ile Lys Lys Gly Leu Asn Lys Leu Ile Asp Gly 900 905 910 Leu Asn Trp Val Gly Gly Lys Leu Gly Met Asp Lys Ile Pro Lys Leu 915 920 925 His Thr Gly Thr Glu His Thr His Thr Thr Thr Arg Leu Val Lys Asn 930 935 940 Gly Lys Ile Ala Arg Asp Thr Phe Ala Thr Val Gly Asp Lys Gly Arg 945 950 955 960 Gly Asn Gly Pro Asn Gly Phe Arg Asn Glu Met Ile Glu Phe Pro Asn 965 970 975 Gly Lys Arg Val Ile Thr Pro Asn Thr Asp Thr Thr Ala Tyr Leu Pro 980 985 990 Lys Gly Ser Lys Val Tyr Asn Gly Ala Gln Thr Tyr Ser Met Leu Asn 995 1000 1005 Gly Thr Leu Pro Arg Phe Ser Leu Gly Thr Met Trp Lys Asp Ile Lys 1010 1015 1020 Ser Gly Ala Ser Ser Ala Phe Asn Trp Thr Lys Asp Lys Ile Gly Lys 1025 1030 1035 1040 Gly Thr Lys Trp Leu Gly Asp Lys Val Gly Asp Val Leu Asp Phe Met 1045 1050 1055 Glu Asn Pro Gly Lys Leu Leu Asn Tyr Ile Leu Glu Ala Phe Gly Ile 1060 1065 1070 Asp Phe Asn Ser Leu Thr Lys Gly Met Gly Ile Ala Gly Asp Ile Thr 1075 1080 1085 Lys Ala Ala Trp Ser Lys Ile Lys Lys Ser Ala Thr Asp Trp Ile Lys 1090 1095 1100 Glu Asn Leu Glu Ala Met Gly Gly Gly Asp Leu Val Gly Gly Ile Leu 1105 1110 1115 1120 Asp Pro Asp Lys Ile Asn Tyr His Tyr Gly Arg Thr Ala Ala Tyr Thr 1125 1130 1135 Ala Ala Thr Gly Arg Pro Phe His Glu Gly Val Asp Phe Pro Phe Val 1140 1145 1150 Tyr Gln Glu Val Arg Thr Pro Met Gly Gly Arg Leu Thr Arg Met Pro 1155 1160 1165 Phe Met Ser Gly Gly Tyr Gly Asn Tyr Val Lys Ile Thr Ser Gly Val 1170 1175 1180 Ile Asp Met Leu Phe Ala His Leu Lys Asn Phe Ser Lys Ser Pro Pro 1185 1190 1195 1200 Ser Gly Thr Met Val Lys Pro Gly Asp Val Val Gly Leu Thr Gly Asn 1205 1210 1215 Thr Gly Phe Ser Thr Gly Pro His Leu His Phe Glu Met Arg Arg Asn 1220 1225 1230 Gly Arg His Phe Asp Pro Glu Pro Tyr Leu Arg Asn Ala Lys Lys Lys 1235 1240 1245 Gly Arg Leu Ser Ile Gly Gly Gly Gly Ala Thr Ser Gly Ser Gly Ala 1250 1255 1260 Thr Tyr Ala Ser Arg Val Ile Arg Gln Ala Gln Ser Ile Leu Gly Gly 1265 1270 1275 1280 Arg Tyr Lys Gly Lys Trp Ile His Asp Gln Met Met Arg Val Ala Lys 1285 1290 1295 Arg Glu Ser Asn Tyr Gln Ser Asn Ala Val Asn Asn Trp Asp Ile Asn 1300 1305 1310 Ala Gln Arg Gly Asp Pro Ser Arg Gly Leu Phe Gln Ile Ile Gly Ser 1315 1320 1325 Thr Phe Arg Ala Asn Ala Lys Arg Gly Tyr Thr Asn Phe Asn Asn Pro 1330 1335 1340 Val His Gln Gly Ile Ser Ala Met Gln Tyr Ile Val Arg Arg Tyr Gly 1345 1350 1355 1360 Trp Gly Gly Phe Lys Arg Ala Gly Asp Tyr Ala Tyr Ala Thr Gly Gly 1365 1370 1375 Lys Val Phe Asp Gly Trp Tyr Asn Leu Gly Glu Asp Gly His Pro Glu 1380 1385 1390 Trp Ile Ile Pro Thr Asp Pro Ala Arg Arg Asn Asp Ala Met Lys Ile 1395 1400 1405 Leu His Tyr Ala Ala Ala Glu Val Arg Gly Lys Lys Ala Ser Lys Asn 1410 1415 1420 Lys Arg Pro Ser Gln Leu Ser Asp Leu Asn Gly Phe Asp Asp Pro Ser 1425 1430 1435 1440 Leu Leu Leu Lys Met Ile Glu Gln Gln Gln Gln Gln Ile Ala Leu Leu 1445 1450 1455 Leu Lys Ile Ala Gln Ser Asn Asp Val Ile Ala Asp Lys Asp Tyr Gln 1460 1465 1470 Pro Ile Ile Asp Glu Tyr Ala Phe Asp Lys Lys Val Asn Ala Ser Ile 1475 1480 1485 Glu Lys Arg Glu Arg Gln Glu Ser Thr Lys Val Lys Phe Arg Lys Gly 1490 1495 1500 Gly Ile Ala Ile Gln 1505 28 261 DNA Staphylococcus bacteriophage CDS (1)..(258) 28 atg tat tac aaa att ggt gag ata aaa aac aaa att ata agc ttt aac 48 Met Tyr Tyr Lys Ile Gly Glu Ile Lys Asn Lys Ile Ile Ser Phe Asn 1 5 10 15 ggg ttt gaa ttt aaa gtg tct gtg atg aag aga cat gac ggt atc agt 96 Gly Phe Glu Phe Lys Val Ser Val Met Lys Arg His Asp Gly Ile Ser 20 25 30 ata caa atc aag gat atg aat aat gtt cca ctt aaa tcg ttt cat gtc 144 Ile Gln Ile Lys Asp Met Asn Asn Val Pro Leu Lys Ser Phe His Val 35 40 45 ata gat tta agc gaa cta tat att gcg acg gat gca atg cgt gac gtt 192 Ile Asp Leu Ser Glu Leu Tyr Ile Ala Thr Asp Ala Met Arg Asp Val 50 55 60 ata aac gaa tgg att gaa aat aac aca gat gaa cag gac aaa cta att 240 Ile Asn Glu Trp Ile Glu Asn Asn Thr Asp Glu Gln Asp Lys Leu Ile 65 70 75 80 aac tta gtc atg aaa tgg tag 261 Asn Leu Val Met Lys Trp 85 29 86 PRT Staphylococcus bacteriophage 29 Met Tyr Tyr Lys Ile Gly Glu Ile Lys Asn Lys Ile Ile Ser Phe Asn 1 5 10 15 Gly Phe Glu Phe Lys Val Ser Val Met Lys Arg His Asp Gly Ile Ser 20 25 30 Ile Gln Ile Lys Asp Met Asn Asn Val Pro Leu Lys Ser Phe His Val 35 40 45 Ile Asp Leu Ser Glu Leu Tyr Ile Ala Thr Asp Ala Met Arg Asp Val 50 55 60 Ile Asn Glu Trp Ile Glu Asn Asn Thr Asp Glu Gln Asp Lys Leu Ile 65 70 75 80 Asn Leu Val Met Lys Trp 85 30 216 DNA Staphylococcus bacteriophage CDS (1)..(213) 30 atg aat ata atg caa ttc aaa agc tta ttg aaa tcg atg tat gaa gag 48 Met Asn Ile Met Gln Phe Lys Ser Leu Leu Lys Ser Met Tyr Glu Glu 1 5 10 15 aca aag caa agc gac ccg att gta gca aat gta tat atc gag act ggt 96 Thr Lys Gln Ser Asp Pro Ile Val Ala Asn Val Tyr Ile Glu Thr Gly 20 25 30 tgg gcg gtc aat aga ttg ttg gac aat aac gag tta tcg cct ttc gat 144 Trp Ala Val Asn Arg Leu Leu Asp Asn Asn Glu Leu Ser Pro Phe Asp 35 40 45 gat tac gac aga gtt gaa aag aaa atc atg aat gaa atc aac tgg aag 192 Asp Tyr Asp Arg Val Glu Lys Lys Ile Met Asn Glu Ile Asn Trp Lys 50 55 60 aaa aca cac att aag gag tgt taa 216 Lys Thr His Ile Lys Glu Cys 65 70 31 71 PRT Staphylococcus bacteriophage 31 Met Asn Ile Met Gln Phe Lys Ser Leu Leu Lys Ser Met Tyr Glu Glu 1 5 10 15 Thr Lys Gln Ser Asp Pro Ile Val Ala Asn Val Tyr Ile Glu Thr Gly 20 25 30 Trp Ala Val Asn Arg Leu Leu Asp Asn Asn Glu Leu Ser Pro Phe Asp 35 40 45 Asp Tyr Asp Arg Val Glu Lys Lys Ile Met Asn Glu Ile Asn Trp Lys 50 55 60 Lys Thr His Ile Lys Glu Cys 65 70 32 186 DNA Staphylococcus bacteriophage CDS (1)..(183) 32 atg caa caa caa gca tat ata aac gca aca att gat ata aga ata cct 48 Met Gln Gln Gln Ala Tyr Ile Asn Ala Thr Ile Asp Ile Arg Ile Pro 1 5 10 15 aca gaa gtt gaa tat cag cat tac gat gat gtg gat aaa gaa aaa gat 96 Thr Glu Val Glu Tyr Gln His Tyr Asp Asp Val Asp Lys Glu Lys Asp 20 25 30 acg ctg gca aag cgc tta gat gac aat ccg gac gaa tta cta aag tat 144 Thr Leu Ala Lys Arg Leu Asp Asp Asn Pro Asp Glu Leu Leu Lys Tyr 35 40 45 gac aac ata aca ata aga cat gca tat ata gag gtg gaa taa 186 Asp Asn Ile Thr Ile Arg His Ala Tyr Ile Glu Val Glu 50 55 60 33 61 PRT Staphylococcus bacteriophage 33 Met Gln Gln Gln Ala Tyr Ile Asn Ala Thr Ile Asp Ile Arg Ile Pro 1 5 10 15 Thr Glu Val Glu Tyr Gln His Tyr Asp Asp Val Asp Lys Glu Lys Asp 20 25 30 Thr Leu Ala Lys Arg Leu Asp Asp Asn Pro Asp Glu Leu Leu Lys Tyr 35 40 45 Asp Asn Ile Thr Ile Arg His Ala Tyr Ile Glu Val Glu 50 55 60 34 86 PRT Staphylococcus bacteriophage 34 Met Tyr Tyr Glu Ile Gly Glu Ile Ile Arg Lys Asn Ile His Val Asn 1 5 10 15 Gly Phe Asp Phe Lys Leu Phe Ile Leu Lys Gly His Met Gly Ile Ser 20 25 30 Ile Gln Val Lys Asp Met Asn Asn Val Pro Ile Lys His Ala Tyr Val 35 40 45 Val Asp Glu Asn Asp Leu Asp Met Ala Ser Asp Leu Phe Asn Gln Ala 50 55 60 Ile Asp Glu Trp Ile Glu Glu Asn Thr Asp Glu Gln Asp Arg Leu Ile 65 70 75 80 Asn Leu Val Met Lys Trp 85 35 98 PRT Staphylococcus bacteriophage 35 Met Phe Asn Ile Lys Arg Lys Thr Glu Glu Val Lys Met Tyr Tyr Glu 1 5 10 15 Ile Gly Glu Ile Ile Arg Lys Asn Ile His Val Asn Gly Phe Asp Phe 20 25 30 Lys Leu Phe Ile Leu Lys Gly His Met Gly Ile Ser Ile Gln Val Lys 35 40 45 Asp Met Asn Asn Val Pro Ile Lys His Ala Tyr Val Val Asp Glu Asn 50 55 60 Asp Leu Asp Met Ala Ser Asp Leu Phe Asn Gln Ala Ile Asp Glu Trp 65 70 75 80 Ile Glu Glu Asn Thr Asp Glu Gln Asp Arg Leu Ile Asn Leu Val Met 85 90 95 Lys Trp 36 10 DNA Artificial Sequence Description of Artificial Sequence Synthetic Sal I restriction site 36 gcgtcgaccg 10 

What is claimed is:
 1. An isolated, purified, or enriched polypeptide comprising at least a fragment of a protein encoded by Staphylococcus aureus bacteriophage 3A open reading frame 33, 41 or 79, a bacteriophage 77 open reading frame 1, or a bacteriophage 96 open reading frame 48, 78 or 100, wherein said fragment is at least 15, contiguous amino acid residues in length.
 2. The polypeptide of claim 1, wherein said polypeptide comprises a fragment at least 30 amino acid residues in length of a said polypeptide normally encoded by said bacteriophage.
 3. A novel protein which is encoded by a nucleic acid molecule which corresponds to a nucleic acid molecule from Staphylococcus aureus bacteriophages 3A, 77 or 96, as shown in SEQ ID Nos.: 17, 18 and 19, respectively.
 4. The novel protein of claim 3, wherein said protein is isolated from a bacteriophage.
 5. The polypeptide of claim 2, wherein said polypeptide comprises a fragment at least 50 amino acid residues in length of a said polypeptide normally encoded by said bacteriophage.
 6. The polypeptide of claim 1, wherein said fragment binds to a bacterial polypeptide bound by a full-length protein encoded by said Staphylococcus aureus bacteriophage open reading frame.
 7. The polypeptide of claim 1, wherein said polypeptide has the amino acid sequence of SEQ ID NO: 31 (96ORF78).
 8. The novel protein of claim 3, wherein said protein is encoded by a nucleic acid molecule having at least 50% identity with nucleic acids 10148 to 10363 of SEQ ID NO:
 19. 9. An isolated, purified, or enriched polypeptide having at least 50% identity with the amino acid sequence of SEQ ID NO: 31 (96ORF78).
 10. The polypeptide of claim 9, wherein said identity is at least 75%.
 11. The polypeptide of claim 10, wherein said identity is at least 95%. 